Team:Harvard/Plasmids

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(New page: == Vectors == *pACYC-Duet vector map *[http://partsregistry.org/wiki/index.php?title=Part:pSB3K3 pSB3K3] ==Parts== *[http://spreadsheets.google.com/ccc?key=pwvx...)
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== Vectors ==
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*[[Media:PACYC-map.pdf‎|pACYC-Duet vector map]]
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*[http://partsregistry.org/wiki/index.php?title=Part:pSB3K3 pSB3K3]
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==Parts==
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<html><a href = "https://2008.igem.org/Team:Harvard"><img src="https://static.igem.org/mediawiki/2008/b/b9/Harvard_logo.png"></a></html>
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{|align="justify" style="background-color:#FFFFFF;text-indent: 15pt;text-align:justify" cellpadding="50" width="90%"
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=Plasmids=
*[http://spreadsheets.google.com/ccc?key=pwvxAM8xwIq37aJ-qKhmUag&hl=en Google Spreadsheet]
*[http://spreadsheets.google.com/ccc?key=pwvxAM8xwIq37aJ-qKhmUag&hl=en Google Spreadsheet]
-
{| {{table}}
+
{|
-
| align="center" style="background:#eeeeee;"|'''Name'''
+
| align="center" style="background:#f0f0f0;"|'''Internal name'''
-
| align="center" style="background:#eeeeee;"|'''Registry Name'''
+
| align="center" style="background:#f0f0f0;"|'''Description'''
-
| align="center" style="background:#eeeeee;"|'''Description'''
+
| align="center" style="background:#f0f0f0;"|'''Registry Name'''
-
| align="center" style="background:#eeeeee;"|'''Origin'''
+
| align="center" style="background:#f0f0f0;"|'''Part Size'''
-
| align="center" style="background:#eeeeee;"|'''Size'''
+
| align="center" style="background:#f0f0f0;"|'''Vector (size)'''
-
| align="center" style="background:#eeeeee;"|'''Marker'''
+
| align="center" style="background:#f0f0f0;"|'''Plasmid size (vector + part)'''
-
| align="center" style="background:#eeeeee;"|'''Transformed?'''
+
| align="center" style="background:#f0f0f0;"|'''Origin'''
-
| align="center" style="background:#eeeeee;"|'''Miniprepped?'''
+
| align="center" style="background:#f0f0f0;"|'''Marker'''
-
| align="center" style="background:#eeeeee;"|'''Culture in Glycerol Stock?'''
+
| align="center" style="background:#f0f0f0;"|'''Transformed?'''
-
| align="center" style="background:#eeeeee;"|'''Sequenced?'''
+
| align="center" style="background:#f0f0f0;"|'''Miniprep'''
 +
| align="center" style="background:#f0f0f0;"|'''Glycerol Stock'''
 +
| align="center" style="background:#f0f0f0;"|'''Sequenced?'''
|-
|-
-
| P1 || [http://partsregistry.org/Part:BBa_I20259 J23113/I20259]||Low promoter GFP tester||p15A pMR101-derived||919||Kan||in S1 and TOP10||Yes for both strains|| Yes for both strains.||||
+
| P1 ||Low promoter+RBS +GFP+term||J23113/I20259||919||pSB3K3 (2750)||3669||p15A||Kan||in S1 and TOP10||Yes for both strains||Yes for both strains.||S1P1A_A, P1E2 (tube labeled E1A)
|-
|-
-
| P2 || [http://partsregistry.org/Part:BBa_I20269 J23150/I20269]||Medium promoter GFP tester||p15A pMR101-derived||919||Kan||in TOP10, not S1||||
+
| P2 ||Med promoter+RBS+ GFP+term||J23150/I20269||919||pSB3K3 (2750)||3669||p15A||Kan||in TOP10, not S1||Yes||||P2E2 (tube labeled 2B)
|-
|-
-
| P3 || [http://partsregistry.org/Part:BBa_I20270 J23151/I20270]||High promoter GFP tester||p15A pMR101-derived||919||Kan||in TOP10 and S1||Yes for both strains.||Yes for both strains.||
+
| P3 ||High promoter+RBS+ GFP+term||J23151/I20270||919||pSB3K3 (2750)||3669||p15A||Kan||in TOP10 and S1||Yes for both strains.||Yes for both strains.||P3E2 (tube labeled E3B), S1P3A_A
|-
|-
-
| P4 || [http://www.merckbiosciences.co.uk/html/NVG/duettable.html pACYC duet]||Low-Medium copy vector||p15a||4008||Cm||Yes||in S1 and TOP10||||
+
| P4 ||Low-Medium copy vector||pACYC duet||4008||||||p15a||Cm||in S1 and TOP10||Yes, for both strains.||Yes for both strains.||
|-
|-
-
| P5 || [http://partsregistry.org/Part:pSB3K3 pSB3K3]||Low-Medium copy vector (w/ death gene)||p15a||2750||Kan||||||
+
| P5 ||Low-Medium copy vector (w/ death gene)||pSB3K3||2750||||||p15a||Kan||in E3||Yes||Yes (B- done 6/30. A-done 7/1)||
|-
|-
-
| P6 || [http://partsregistry.org/Part:BBa-E1010 BBa-E1010]||RFP only (w/ death gene)||pMB1||681||Kan||||||
+
| P11 ||Light responsive system, dual regulation||BBa_M30109||4333||pSB1AC3 (3055)||7388||pMB1||Amp/Cm||Failed||||||
|-
|-
-
| P7 || [http://partsregistry.org/Part:pSB1A2 pSB1A2]||GFP only||pMB1||2079||Amp||||||
+
| P13 ||Low copy plasmid||pCDFDuet-1||3781||||||CloDF13||Sm||in S1||Yes, 435 ng/uL||Yes||
|-
|-
-
| P8 || [http://partsregistry.org/Part:BBa_J04450 BBa_J04450]||RFP with LacI promoter||pMB1||1069||Amp||||||
+
| P15 ||pTet+RBS+GFP+term||BBa_I13522||937||BBa_J23006 (3279)||||ColE1||Amp||in E2||Yes||No||P15A, P15B
|-
|-
-
| P9 || [http://partsregistry.org/Part:BBa_J04430 BBa_J04430]||GFP with LacI promoter||pMB1||1083||Amp||||||
+
| P16 ||Tet repressible promoter||BBa_R0040||54||||||pMB1||Amp||in E2||Yes||No||
|-
|-
-
| P10 ||[http://partsregistry.org/Part:BBa_I715038 BBa_I715038]||T7 Polymerase with LacI promoter||pMB1||2878||Amp||||||
+
| P17 ||Inverter (TetR w/o promoter and Tet promoter)||BBa_Q04400||902||pSB2K3 (4425)||5327||OriS, P1 lytic, F1||Kan||in E1||BAD||BAD||cells from MIT correct
|-
|-
-
| P11 || [http://partsregistry.org/Part:BBa_M30109 BBa_M30109]||Light responsive system, dual regulation||pUC19-derived pMB1||4333||Amp/Cm||||||
+
| P18 ||cI regulated promoter||R0051||49||||||pMB1||Amp||in E2||Yes||Yes, 7/4||P18A, P18B appear so (20 bp align at end, but ref is 49 bp, so this isn\'t bad)
|-
|-
-
| P12 || [http://www.merckbiosciences.co.uk/html/NVG/duettable.html pETDuet-1]||Low copy plasmid||pBR322-derived ColE1||5420||Amp||||||
+
| P20 ||GFP (LVA tagged) with Lac promoter||BBa_J04431||1122||||||pMB1||Amp, Kan||in E2||Yes||No||?! P20A: ~100 bp of beginning of F read aligned to ref; R read did not
|-
|-
-
| P13 || [http://www.merckbiosciences.co.uk/html/NVG/duettable.html pCDFDuet-1]||Low copy plasmid||CloDF13||3781||Sm||Yes||in S1||
+
| P21 ||pACYC (P4) w/ biobricks||A1 (pACYC)||???||||||???||???||in S1 and TOP10||Yes||Yes, for both strains.||
|-
|-
-
| P14 || [http://www.merckbiosciences.co.uk/html/NVG/duettable.html pCOLADuet-1]||Low copy plasmid||ColA||3719||Kan||||||
+
| P22 ||ompR controlled promoter for ompC (long)||BBa_R0082||108||||||pMB1||Amp||||||||WRONGWRONGWRONG: DID NOT ALIGN TO REF
|-
|-
-
| P15 || [http://partsregistry.org/Part:BBa_I13522 BBa_I13522]||GFP with Tet promoter||ColE1||937||Amp||||||
+
| P23 ||ompR controlled promoter for ompC (short)||BBa_R0083||78||||||pMB1||Amp||in E2||Yes||No||WRONGWRONGWRONG: DID NOT ALIGN TO REF
|-
|-
-
| P16 || [http://partsregistry.org/Part:BBa_R0040 BBa_R0040]||Tet repressible promoter||pMB1||54||Amp||||||
+
| P24 ||ompR controlled promoter for ompF||BBa_R0084||108||||||pMB1||Amp||in E2||Yes||No||WRONGWRONGWRONG: DID NOT ALIGN TO REF
|-
|-
-
| P17 || [http://partsregistry.org/Part:BBa_Q04400 BBa_Q04400]||Inverter (TetR w/o promoter and Tet promoter)||OriS, P1 lytic, F1||902||Kan||||||
+
| P25 ||Promoter activated by OmpR-P with the reporter GFP||BBa_J58102||991||||||pMB1||Amp||||||||WRONGWRONGWRONG: DID NOT ALIGN TO REF
|-
|-
-
| P18 || [http://partsregistry.org/Part:BBa_R0051 BBa_R0051]||lambda promoter (cI regulated)||pMB1||49||Amp||||||
+
| P26 ||Mutated promoter activated by OmpR-P with the reporter GFP||BBa_J58103||961||||||pMB1||Amp||in E2||Yes||No||WRONGWRONGWRONG: DID NOT ALIGN TO REF
|-
|-
-
| P19 || [http://partsregistry.org/Part:BBa_I763007 BBa_I763007]||RFP with lambda promoter||pMB1||918||Amp||||||
+
| P27 ||repressor plasmid w/ constitutive expression of LacI and TetR||pZS4int2||||||||pSC101||Sm||Yes||Yes (6/26)||Yes (6/26)||
|-
|-
-
| P20 || [http://partsregistry.org/Part:BBa_J04431 BBa_J04431]||GFP (LVA tagged) with Lac promoter||pMB1||1122||Amp, Kan||||||
+
| P28 ||GFP w/ PLtctO-1||pZE21||||||||ColE1||Kan||Yes||Yes (6/26)||Yes (6/26)||
|-
|-
-
| P21 ||A1 (pACYC)||pACYC (P4) w/ biobricks||???||???||???||Yes||in S1 and TOP10||
+
| P29 ||Venus (mutant YFP) PL (Plambda)||pZS*2R||||||||pSC101*||Kan||Yes||Yes (6/26)||Yes (6/26)||
|-
|-
-
| P22 || [http://partsregistry.org/Part:BBa_R0082 BBa_R0082]||ompR controlled promoter for ompC (long)||pMB1||108||Amp||||||
+
| P30 ||YFP w/ PLlacO-1||pZA32||||||||p15A||Cm||Yes||Yes (6/26)||Yes (6/26)||
|-
|-
-
| P23 || [http://partsregistry.org/Part:BBa_R0083 BBa_R0083]||ompR controlled promoter for ompC (short)||pMB1||78||Amp||||||
+
| P31 ||Venus w/ PLlacO-1||pZS*1R||||||||pSC101*||Amp||Yes||Yes (6/26)||Yes (6/26)||
|-
|-
-
| P24 || [http://partsregistry.org/Part:BBa_R0084 BBa_R0084]||ompR controlled promoter for ompF||pMB1||108||Amp||||||
+
| P32 ||Venus w/ PLlacO-1||pZA32||||||||p15A||Cm||Yes||Yes (6/26)||Yes (6/26)||
|-
|-
-
| P25 || [http://partsregistry.org/Part:BBa_J58102 BBa_J58102]||Promoter activated by OmpR-P with the reporter GFP||pUC19-derived pMB1||991||Amp||||||
+
| P33||Venus w/ PLtctO-1||pZA31||||||||p15A||Cm||Yes||Yes (6/26)||Yes (6/26)||
|-
|-
-
| P26 || [http://partsregistry.org/Part:BBa_J58103 BBa_J58103]||Mutated promoter activated by OmpR-P with the reporter GFP||pUC19-derived pMB1||961||Amp||||||
+
| P34 ||GFP w/ PLlacO-1||pZE12||||||||ColE1||Amp||Yes||Yes (6/26)||Yes (6/26)||
|-
|-
-
| P27 ||pZS4int2||repressor plasmid||pSC101||||Sm||Yes||||
+
| P35 ||accessory ||pLDR8||||||||||||Yes||Yes (6/26)||Yes (6/26)||
|-
|-
-
| P28 ||pZE21||GFP||ColE1||||Kan||Yes||||
+
| P36 ||same as P20||BBa_I763004||||||||||Kan/Amp||||||||
|-
|-
-
| P29 ||pZS*2R||Venus (mutant YFP)||pSC101*||||Kan||Yes||||
+
| P37 ||Constructive expression LacI w/ pTetR promoter||BBa_I722007||||||||||Amp||Failed (liquid cultures didn\'t grow)||||||
|-
|-
-
| P30 ||pZA32||YFP||p15A||||Cm||Yes||||
+
| P38 ||High constitutive promoter||BBa_J23114||35||||||||Amp||2007 in E1 (DH5a), 2008 Failed, MIT||Yes BUT funny digestion pattern||2007 (B- done 6/30. A- done 7/1)||
|-
|-
-
| P31 ||pZS*1R||Venus||pSC101*||||Amp||Yes||||
+
| P39 ||Low constitutive promoter||BBa_J23113||35||||||||Amp||2008 (A grew), 2007 (A grew), MIT||Yes BUT funny digestion pattern||2008 (A- done 6/30) 2007 (A-done 6/30)||
|-
|-
-
| P32 ||pZA32||Venus||p15A||||Cm||Yes||||
+
| P40 ||Ribosome Binding Site||BBa_B0032||13||||||||Amp||2008 (A grew), 2007 (B grew)||Yes.||2007 (B -done 6/30) 2008 (A-done 6/30)||
|-
|-
-
| P33 ||pZA31||Venus||p15A||||Cm||Yes||||
+
| P41 ||Transcriptional Terminator||BBa_B0010||||||||||Amp||2007 (A and B grew, 7/2), 2008 (B grew, 7/2)||||2007 (A and B done 7/4), 2008 (B done 7/4)||
|-
|-
-
| P34 ||pZE12||GFP||ColE1||||Amp||Yes||||
+
| P42 ||LacI coding region||\"BBa_C0012
|-
|-
-
| P35 ||pLDR8||||||||||Yes||||
+
| \"||1128||||||||Amp||Failed (liquid cultures didn\'t grow)||||||
|-
|-
-
| P36 || [http://partsregistry.org/Part:BBa_I763004 BBa_I763004]||GFP + IPTG + LVA||||||Kan/Amp||||||
+
| P43 ||TetR coding region||BBa_C0040||660||||||||Amp||2007 (A grew)||Yes||2007 (A-done 6/30)||P43A E1 07
|-
|-
-
| P37 || [http://partsregistry.org/wiki/index.php/Part:BBa_I722007 BBa_I722007]||Constructive expression LacI w/ pTetR promoter||||||Amp||||||
+
| P44 ||cI lambda||\"BBa_C0051
|-
|-
-
| P38 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23114 BBa_J23114]||High constitutive promoter||||||Amp||||||
+
| \"||750||||||||Amp||2007 (A grew)||Yes||2007 (A-done 6/30)||P44A E1 07
|-
|-
-
| P39 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23113 BBa_J23113]||Low constitutive promoter||||||Amp||||||
+
| P45 ||GFP only w/ RBS & terminator||BBa_E0240||876||pSB1A2 (2079)||2955||pMB1||Amp||2007 (A and B grew)||Yes||2007 (A and B -done 6/30)||P45A, P45B >E1 07 for both
|-
|-
-
| P40 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0032 BBa_B0032]||Ribosome Binding Site||||||Amp||||||
+
| P46 ||Base Vector||BBa_I51020||||||||||Amp||A and B grew.||Yes||A and B -done 6/30||
|-
|-
-
| P41 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0010 BBa_B0010]||Transcriptional Terminator||||||Amp||||||
+
| P47 ||Kan Resistance Cassette||BBa_P1003||967||||||||Kan||2007 (A and B grew, but spilled)||Yes||2007 (A and B -done 7/1)||P47A, P47B >E1 07 for both
|-
|-
-
| P42 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0012 BBa_C0012]||LacI coding region||||||Amp||||||
+
| P48 ||Cm Resistance Cassette||BBa_P1004||769||pSB1A1 (2708)||3477||||Cm||2007 (A and B grew, but spilled)||Yes||2007 (A and B -done 7/1).  Duplicate done 7/4.||P48B E1 07
|-
|-
-
| P43 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0040]||TetR coding region||||||Amp||||||
+
| P51||LacI QPI w/ RBS & promoter||BBa_Q04121||1370||pSB2K3 (4425)||5795||pMB1||Kan||||||||cells from MIT correct
|-
|-
-
| P44 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0051 BBa_C0051]||cI lambda||||||Amp||||||
+
| P52||cI QPI w/ RBS & promoter||BBa_Q04510||987||pSB2K3 (4425)||5412||pMB1||Kan||||||||
|-
|-
-
| P45 || [http://partsregistry.org/wiki/index.php?title=Part:BBa_E0240 BBa_E0240]||GFP only w/ RBS & terminator||||||Amp||||||
+
| P53||PoPS --> TetR||BBa_P0440||840||||||||Amp||2007 (B grew), 2008 (A grew)||Yes. (7/3).||2007 (B done 7/3), 2008 (A done 7/3)||
|-
|-
-
| P46 || [http://partsregistry.org/Part:BBa_I51020 BBa_I51020]||Base Vector|||||||Amp||||||
+
| P54||PoPS --> LacI||BBa_P0412||1308||||||||Amp||2007 (A grew)||Yes. (7/3).||2007 (A done 7/3)||
|-
|-
-
| P47 || [http://partsregistry.org/Part:BBa_P1003 BBa_P1003]||Kan Resistance Cassette||||||Kan||||||
+
| P55||RBS.Lambda.cI. LVA.Terminator||BBa_P0455||896||||||||A/C||2007 (A and B grew). 2008 (A grew)||Yes for A and B (7/3).||2007 (A and B done 7/3). 2008 (A grew)||
|-
|-
-
| P48 || [http://partsregistry.org/Part:BBa_P1004 BBa_P1004]||Cm Resistance Cassette||||||Cm||||||
+
| P56||||RBS+TetR coding region (P40 and P43)||||||||||Amp||Yes in E1.||Yes for A and B (7/3).||A and B done 7/3||
|-
|-
-
| P49 || [http://partsregistry.org/Part:BBa_P1002 BBa_P1002]||Amp Resistance Cassette|||||||Amp||||||
+
| P57||RBS+lambda cI coding region (P40 and P44)||Composite||||||||||Amp||Yes in E1.||Yes for A and B (7/3).||A and B done 7/3||
|-
|-
-
| P50 || [http://partsregistry.org/Part:BBa_P1005 BBa_P1005]||Tet Resistance Cassette||||||Tet||||||
+
| P58||GFP under pTet (with RBS and term) on plasmid with p15A origin (P1 vector + P15 part)||Composite||3016||||||||Kan||Yes in E1.||Yes for A and B (7/3).||A and B done 7/3||
|-
|-
-
| P51 || [http://partsregistry.org/Part:BBa_Q04121 BBa_Q04121]||LacI QPI w/ RBS & promoter||||||Kan||||||
+
| P59||GFP under pLac (with RBS and term) on plasmid with p15A origin (P1 vector + P20 part)||Composite||3201||||||||Kan||Yes in E1.||Yes for A and B (7/3).||A and B done 7/3||
|-
|-
-
| P52 || [http://partsregistry.org/Part:BBa_Q04510 BBa_Q04510]||cI Lambda QPI||||||Kan||||||
+
| P60||||Composite ||||||||||||||||||
|-
|-
-
| P53 || [http://partsregistry.org/Part:BBa_P0440 BBa_P0440]||PoPS -> TetR||||||Amp||||||
+
| P61||Terminator||BBa_B0011||||||||||||2008 (A grew.)||Yes for 2008 A.||2008 (A done 7/4)||
|-
|-
-
| P54 || [http://partsregistry.org/Part:BBa_P0412 BBa_P0412]||PoPS -> LacI||||||Amp||||||
+
| P62||Terminator||BBa_B0012||||||||||||2007 (A and B grew). 2008 ((Plate yielded 1 colony only- failed to grow)||Yes for A and B 2007.||2007 (A and B done 7/4)||
|-
|-
-
| P55 || [http://partsregistry.org/Part:BBa_P0455 BBa_P0455]||RBS.cI.LVA.Term||||||Amp/Cm||||||
+
| P63||Double terminator (P62 and P61)||BBa_B0014||95||pSB1AK3 (3189)||3284||||Amp/Kan||2007 (A and B grew).  2008 (Plate yielded 1 colony only- failed to grow)||Yes for A and B 2007.||2007 (A and B done 7/4)||
 +
|-
 +
| P64||Double terminator (P60 and P62)||BBa_B0015||||||||||||2007 (A grew)||||2007 (A done 7/4)||
 +
|-
 +
| P65||Double terminator (P62 and P60)||BBa_B0025||||||||||||Failed (liquid cultures didn\'t grow)||||||
 +
|-
 +
| P66||hi promoter+RBS+TetR coding region (P38+P56)||Composite||||||||||||||||||
 +
|-
 +
| P67||Low promoter+RBS+TetR ||Composite||||||||||||||||||
 +
|-
 +
| P68||hi promoter +RBS+lambda cI coding region (P38+P57)||Composite||||||||||||||||||
 +
|-
 +
| P69||Low promoter+RBS+lambda cI repressor||Composite||||||||||||||||||
 +
|-
 +
| P70||P66 and term||Composite||||||||||||||||||
 +
|-
 +
| P71||P67 and term||Composite||||||||||||||||||
 +
|-
 +
| P72||P68 and term||Composite||||||||||||||||||
 +
|-
 +
| P73||P69 and term||Composite||||||||||||||||||
 +
|-
 +
| P74||lambda cI promoter+RBS+GFP+term (P45 + P18)||Composite||||||||||||Yes.||yes||||
 +
|-
 +
| P75||P74 + P1||Composite||||||||||||yes in both S1 and E1||yes 7/22||||
 +
|-
 +
| P76||P17+P45||Tet QPI+GFP||1778||pSB1A2 (2079)||3857||pMB1||Amp||Yes||Yes||Yes||
 +
|-
 +
| P77||RBS + LacI + terminators||BBa_P0312||1307||||||||||Failed (liquid cultures didn\'t grow)||||||
 +
|-
 +
| P78||RBS + LacI + terminators||BBa_P0112||1310||||||||||Failed (liquid cultures didn\'t grow)||||||
 +
|-
 +
| P79||P90+ P48||Composite||||||||||||||||||
 +
|-
 +
| P80||P90 + P49||Composite||||||||||||no- wrong||||||
 +
|-
 +
| P83||P45+mtrA (w/ Pst1)||Composite||||||||||||||||||
 +
|-
 +
| P84||heat sensitive Lac- YFP||J06912||2314||pSB1A2 (2079)||||pMB1||Amp||Yes, in E1.||Yes, 7/9||||
 +
|-
 +
| P85||heat sensitive Lac-YFP||J06911||2314||pSB1A2 (2079)||||pMB1||Amp||Yes, in E1.||yes||||
 +
|-
 +
| P86||pPL-PCB from UT Austin||n/a||||||||P15a||Amp||Yes in S1||||||
 +
|-
 +
| P87||pPL-cph1-envZ-pprotet||n/a||||||||ColE1||Cm||No||||||
 +
|-
 +
| P88||light-sensing EnvZ||BBa_I15010||2238||pSB2K3 (4425)||||F\', P1 lytic||Kan||failed||||||
 +
|-
 +
| P89||RBS + LacI + terminators + pLac||BBa_Q01121||1372||pSB1A2 (2079)||3451||pMB1||Kan||failed||||||
 +
|-
 +
| P90||CDF in P1 vector||||||pSB3K3 (2750)||||p15A||Kan||||||||
 +
|-
 +
| P91||P89+P45: lac QPI+complete GFP||||4327||||||||Amp||||||||
 +
|-
 +
| P92||P84 + P1||Composite||||||||||||Q04510||||||
 +
|-
 +
| P93||P85+P1||Composite||||||||||||||||||
 +
|-
 +
| P94||P40 + mtrB||Composite||>4000||pSB1A2 (2079)||||||Amp||||||||
 +
|-
 +
| P95||RBS + cph8||BBa_S03419||||||||||||||Yes||||
 +
|-
 +
| P96||RBS + cph8 + terminators||BBa_S03417||||||||||||||Yes||||
 +
|-
 +
| P97||Strong RBS||BBa_B0034||12||pSB1A2 (2079)||2.1 kb||pMB1||Amp||Yes||Yes||||
 +
|-
 +
| P99||cell death||BBa_P1010||||||||||||||||||Part from MIT aligned to ref
 +
|-
 +
| P101||pSB4C5 (with death gene)||pSB4C5/I52002||1090||pSB4C5 (3221)||4311||pSC101||Cm||Yes||Yes||Yes||Part from MIT aligned to ref
 +
|-
 +
| P102||P108+P45 (lac induced GFP) in p15a vector||||2281||pSB3K3 (2750)||5031||P15A||Kan||Yes||Yes||Yes||Yes, along with 108+45#41
 +
|-
 +
| P103||P108+P45 (lac induced GFP) in pSC101 vector||||2281||pSB4C5 (3221)||5502||pSC101||Cm||Yes||Yes||Yes||P103 # 41, 45
 +
|-
 +
| P104||P75 + P63||||||||||||||Yes 1 colony||yes 7/28||||
 +
|-
 +
| P105||P87 w/o PstI site (mutated Cph)||||2235||2022||4257||ColE1||Cm||yes||||||E2P105A,  E5P105D
 +
|-
 +
| p106||pER21 - plasmid for genetic recombination||n/a : from Dad||||||||||Gentamycin||Yes||Yes||||
 +
|-
 +
| p107||pKOV - plasmid for genetic recombination||n/a: from George Church||||||||||Cm||Yes||Yes||||
 +
|-
 +
| P108||P3 + (P39+51) beta (lac QPI)||composite||1405||pSB3K3 (2750)||4155||p15A||Kan||yes||yes||yes||Verified
 +
|-
 +
| P109||RBS+full lacZ||I732017||3093||pSB1A2 (2079)||5172||pMB1||Amp||yes||yes||yes||correct from MIT
 +
|-
 +
| P110||RBS+lacZalpha+term||E0435||534||pSB4K5 (3419)||3953||pSC101||Kan||yes||yes||yes||\"correct from MIT
 +
|-
 +
| \"
 +
|-
 +
| P111||RBS+lacZalpha+term||E0435||534||pSB4C5 (3221)||3755||pSC101||Cm||yes||yes||yes||correct from MIT
 +
|-
 +
| P112||RBS+lacZalpha+term||E0435||534||pSB1AK3 ( 3189)||3723||pMB1||A/K||yes||yes||yes||correct from MIT
 +
|-
 +
| P113||RBS+lacZalpha+term||E0435||534||pSB4A3 (3339)||3873||repA pSC101||Amp||yes||yes||yes||correct from MIT
 +
|-
 +
| P114||pJBC2 (inducible IPTG system)||||||||||||||||Yes||||
 +
|-
 +
| P115||low promoter+RBS+E. coli ompR+term (from alt method)||||960||pSB3K3 (2750)||3710||P15A||Kan||Yes||Yes||||ompR+P1 #64, 73 correct
 +
|-
 +
| P116||P38+P17 in pSB3K3||||937||pSB3K3 (2750)||3687||P15A||Kan||Yes||Yes||||P38+17 in P1/3 #2 , 3, 5
 +
|-
 +
| P117||P39+P17 in pSB3K3||||937||pSB3K3 (2750)||3687||P15A||Kan||Yes||Yes||||P39+17 in P1/3 #1
 +
|-
 +
| P118||low promoter+RBS+HO-pcyA+term in pSB3K3||||1555||pSB3K3 (2750)||4305||P15A||Kan||Yes||Yes||||HO+P1 #3
 +
|-
 +
| P119||P63+P18||Composite||||||||||||||||||
 +
|-
 +
| P120||P63 + P18 + cI (RBS primer set)||Composite||||||||||||||||||verified (8/13)
 +
|-
 +
| P121||P104 + cI (RBS primer set)||||||||||||||||||||verified (8/13)
 +
|-
 +
| P122||P120 + P38||||||||||||||||||||
 +
|-
 +
| P123||P120 + P39||||||||||||||||||||
 +
|-
 +
| P124||P121 + P38||||||||||||||||||||
 +
|-
 +
| P125||P121 + P39||||||||||||||||||||
 +
|-
 +
| P126||P115 (low ompR) in pSB4C5||||960||pSB4C5 (3221)||4181||pSC101||Cm||Yes||Yes||Yes||Yes, along with 101+115 numbers 4, 14, 25
 +
|-
 +
| P127||P118 (low HO-pcyA) in pSB4C5||||1555||pSB4C5 (3221)||4776||pSC101||Cm||Yes||Yes||Yes||Yes, along with 101+118 numbers 61, 77
 +
|-
 +
| P128||P97+mtrB||||2106||pSB1A2 (2079)||4185||pMB1||Amp||Yes||Yes||Yes||128# 1, 5, 9, appear to have a silent SNP
 +
|-
 +
| P129||mtrB+P63||||2189||pSB1AK3 (3189)||5378||||Amp/Kan||Yes||Yes||Yes||129# 21, 28 appear to have a silent SNP
 +
|-
 +
| P130||P97+mtrB+P63||Composite||2201||pSB1AK3 (3189)||5390||||Amp/Kan||Yes||||||
 +
|-
 +
| P131||P130 in pSB4C5||||2201||pSB4C5 (3221)||5422||pSC101||Cm||||||||A and B have F>S
 +
|-
 +
| P132||P108 in pSB4C5||||1405||pSB4C5 (3221)||4626||pSC101||Cm||||||||A, D correct
 +
|-
 +
| P133||P116 in pSB4C5||||937||pSB4C5 (3221)||4158||pSC101||Cm||||||||A, B correct
 +
|-
 +
| P134||P117 in pSB4C5||||937||pSB4C5 (3221)||4158||pSC101||Cm||||||||A, B correct
 +
|-
 +
| P135||P39+P130||||2236||pSB1A2 (2079)||4315||pMB1||Amp||||||||#31, 39, 60, 70, 80 F>S, but need more checking for SNPs in mtrB
 +
|-
 +
| P136||P38+P130||||2236||pSB1A2 (2079)||4315||pMB1||Amp||||||||#20 F>S, but needs checking for SNPs in mtrB
 +
|-
 +
| P137||P135 in pSB4C5||||2236||pSB4C5 (3221)||5457||pSC101||Cm||||||||#32- HAS MUTATION IN MIDDLE, F>S
 +
|-
 +
| P138||P136 in pSB4C5||||||||||||||||||||#1, 2> check for mutations in mtrB, are they not on the same vector?
 +
|-
 +
| P139||P130 in P133 (high constitutive promoter+tet QPI+RBS+mtrB+terminator)||||||||||||||no||||||
 +
|-
 +
| P140||P108 in P131 (low constitutive promoter+lac QPI+RBS+mtrB+terminator||||3637 (definite)||pSB4C5 (3221)||6858||pSC101||Cm||||||||YES, but there is a F>S change in mtrB
 +
|-
 +
| P141||P38+51 (high promoter+lac QPI)||||1405||pSB1A2 (2079)||3484||pMB1||Amp||||||||#3, 4 correct
 +
|-
 +
| P142||P141 in P131 (high promoter+lac QPI+RBS+mtrB+terminator)||||3637||pSB4C5 (3221)||6858||pSC101||Cm||||||||YES but needs scrutiny (can\'t check that hi promoter is there, and SNPs in mtrB)
 +
|-
 +
| P143||P130 in P134 (low promoter+tet QPI+RBS+mtrB+term)||||||||||||||no||||||
 +
|-
 +
| P144||high mtrB in 15a||38+130+1||2236||pSB3K3 (2750)||4986||p15A||Kan||yes||Yes||Yes||
 +
|-
 +
| P145||low mtrB in 15a||39+130+1||2236||pSB3K3 (2750)||4986||p15A||Kan||yes||Yes||Yes||
 +
|-
 +
| P146||high lacQPI+ mtrB in 15a||38+51+130+1||3637||pSB3K3 (2750)||6387||p15A||Kan||yes||Yes||Yes||
 +
|-
 +
| P147||low lacQPI+ mtrB in 15a||39+51+130+1||3637||pSB3K3 (2750)||6387||p15A||Kan||yes||Yes||Yes||
 +
|-
 +
|
 +
|}
 +
 
 +
|}
 +
<br><br>
 +
 
 +
<!--- end body --->
|}
|}

Latest revision as of 22:29, 28 October 2008



Plasmids

  • [http://spreadsheets.google.com/ccc?key=pwvxAM8xwIq37aJ-qKhmUag&hl=en Google Spreadsheet]
Internal name Description Registry Name Part Size Vector (size) Plasmid size (vector + part) Origin Marker Transformed? Miniprep Glycerol Stock Sequenced?
P1 Low promoter+RBS +GFP+termJ23113/I20259919pSB3K3 (2750)3669p15AKanin S1 and TOP10Yes for both strainsYes for both strains.S1P1A_A, P1E2 (tube labeled E1A)
P2 Med promoter+RBS+ GFP+termJ23150/I20269919pSB3K3 (2750)3669p15AKanin TOP10, not S1YesP2E2 (tube labeled 2B)
P3 High promoter+RBS+ GFP+termJ23151/I20270919pSB3K3 (2750)3669p15AKanin TOP10 and S1Yes for both strains.Yes for both strains.P3E2 (tube labeled E3B), S1P3A_A
P4 Low-Medium copy vectorpACYC duet4008p15aCmin S1 and TOP10Yes, for both strains.Yes for both strains.
P5 Low-Medium copy vector (w/ death gene)pSB3K32750p15aKanin E3YesYes (B- done 6/30. A-done 7/1)
P11 Light responsive system, dual regulationBBa_M301094333pSB1AC3 (3055)7388pMB1Amp/CmFailed
P13 Low copy plasmidpCDFDuet-13781CloDF13Smin S1Yes, 435 ng/uLYes
P15 pTet+RBS+GFP+termBBa_I13522937BBa_J23006 (3279)ColE1Ampin E2YesNoP15A, P15B
P16 Tet repressible promoterBBa_R004054pMB1Ampin E2YesNo
P17 Inverter (TetR w/o promoter and Tet promoter)BBa_Q04400902pSB2K3 (4425)5327OriS, P1 lytic, F1Kanin E1BADBADcells from MIT correct
P18 cI regulated promoterR005149pMB1Ampin E2YesYes, 7/4P18A, P18B appear so (20 bp align at end, but ref is 49 bp, so this isn\'t bad)
P20 GFP (LVA tagged) with Lac promoterBBa_J044311122pMB1Amp, Kanin E2YesNo?! P20A: ~100 bp of beginning of F read aligned to ref; R read did not
P21 pACYC (P4) w/ biobricksA1 (pACYC)?????????in S1 and TOP10YesYes, for both strains.
P22 ompR controlled promoter for ompC (long)BBa_R0082108pMB1AmpWRONGWRONGWRONG: DID NOT ALIGN TO REF
P23 ompR controlled promoter for ompC (short)BBa_R008378pMB1Ampin E2YesNoWRONGWRONGWRONG: DID NOT ALIGN TO REF
P24 ompR controlled promoter for ompFBBa_R0084108pMB1Ampin E2YesNoWRONGWRONGWRONG: DID NOT ALIGN TO REF
P25 Promoter activated by OmpR-P with the reporter GFPBBa_J58102991pMB1AmpWRONGWRONGWRONG: DID NOT ALIGN TO REF
P26 Mutated promoter activated by OmpR-P with the reporter GFPBBa_J58103961pMB1Ampin E2YesNoWRONGWRONGWRONG: DID NOT ALIGN TO REF
P27 repressor plasmid w/ constitutive expression of LacI and TetRpZS4int2pSC101SmYesYes (6/26)Yes (6/26)
P28 GFP w/ PLtctO-1pZE21ColE1KanYesYes (6/26)Yes (6/26)
P29 Venus (mutant YFP) PL (Plambda)pZS*2RpSC101*KanYesYes (6/26)Yes (6/26)
P30 YFP w/ PLlacO-1pZA32p15ACmYesYes (6/26)Yes (6/26)
P31 Venus w/ PLlacO-1pZS*1RpSC101*AmpYesYes (6/26)Yes (6/26)
P32 Venus w/ PLlacO-1pZA32p15ACmYesYes (6/26)Yes (6/26)
P33Venus w/ PLtctO-1pZA31p15ACmYesYes (6/26)Yes (6/26)
P34 GFP w/ PLlacO-1pZE12ColE1AmpYesYes (6/26)Yes (6/26)
P35 accessory pLDR8YesYes (6/26)Yes (6/26)
P36 same as P20BBa_I763004Kan/Amp
P37 Constructive expression LacI w/ pTetR promoterBBa_I722007AmpFailed (liquid cultures didn\'t grow)
P38 High constitutive promoterBBa_J2311435Amp2007 in E1 (DH5a), 2008 Failed, MITYes BUT funny digestion pattern2007 (B- done 6/30. A- done 7/1)
P39 Low constitutive promoterBBa_J2311335Amp2008 (A grew), 2007 (A grew), MITYes BUT funny digestion pattern2008 (A- done 6/30) 2007 (A-done 6/30)
P40 Ribosome Binding SiteBBa_B003213Amp2008 (A grew), 2007 (B grew)Yes.2007 (B -done 6/30) 2008 (A-done 6/30)
P41 Transcriptional TerminatorBBa_B0010Amp2007 (A and B grew, 7/2), 2008 (B grew, 7/2)2007 (A and B done 7/4), 2008 (B done 7/4)
P42 LacI coding region\"BBa_C0012
\"1128AmpFailed (liquid cultures didn\'t grow)
P43 TetR coding regionBBa_C0040660Amp2007 (A grew)Yes2007 (A-done 6/30)P43A E1 07
P44 cI lambda\"BBa_C0051
\"750Amp2007 (A grew)Yes2007 (A-done 6/30)P44A E1 07
P45 GFP only w/ RBS & terminatorBBa_E0240876pSB1A2 (2079)2955pMB1Amp2007 (A and B grew)Yes2007 (A and B -done 6/30)P45A, P45B >E1 07 for both
P46 Base VectorBBa_I51020AmpA and B grew.YesA and B -done 6/30
P47 Kan Resistance CassetteBBa_P1003967Kan2007 (A and B grew, but spilled)Yes2007 (A and B -done 7/1)P47A, P47B >E1 07 for both
P48 Cm Resistance CassetteBBa_P1004769pSB1A1 (2708)3477Cm2007 (A and B grew, but spilled)Yes2007 (A and B -done 7/1). Duplicate done 7/4.P48B E1 07
P51LacI QPI w/ RBS & promoterBBa_Q041211370pSB2K3 (4425)5795pMB1Kancells from MIT correct
P52cI QPI w/ RBS & promoterBBa_Q04510987pSB2K3 (4425)5412pMB1Kan
P53PoPS --> TetRBBa_P0440840Amp2007 (B grew), 2008 (A grew)Yes. (7/3).2007 (B done 7/3), 2008 (A done 7/3)
P54PoPS --> LacIBBa_P04121308Amp2007 (A grew)Yes. (7/3).2007 (A done 7/3)
P55RBS.Lambda.cI. LVA.TerminatorBBa_P0455896A/C2007 (A and B grew). 2008 (A grew)Yes for A and B (7/3).2007 (A and B done 7/3). 2008 (A grew)
P56RBS+TetR coding region (P40 and P43)AmpYes in E1.Yes for A and B (7/3).A and B done 7/3
P57RBS+lambda cI coding region (P40 and P44)CompositeAmpYes in E1.Yes for A and B (7/3).A and B done 7/3
P58GFP under pTet (with RBS and term) on plasmid with p15A origin (P1 vector + P15 part)Composite3016KanYes in E1.Yes for A and B (7/3).A and B done 7/3
P59GFP under pLac (with RBS and term) on plasmid with p15A origin (P1 vector + P20 part)Composite3201KanYes in E1.Yes for A and B (7/3).A and B done 7/3
P60Composite
P61TerminatorBBa_B00112008 (A grew.)Yes for 2008 A.2008 (A done 7/4)
P62TerminatorBBa_B00122007 (A and B grew). 2008 ((Plate yielded 1 colony only- failed to grow)Yes for A and B 2007.2007 (A and B done 7/4)
P63Double terminator (P62 and P61)BBa_B001495pSB1AK3 (3189)3284Amp/Kan2007 (A and B grew). 2008 (Plate yielded 1 colony only- failed to grow)Yes for A and B 2007.2007 (A and B done 7/4)
P64Double terminator (P60 and P62)BBa_B00152007 (A grew)2007 (A done 7/4)
P65Double terminator (P62 and P60)BBa_B0025Failed (liquid cultures didn\'t grow)
P66hi promoter+RBS+TetR coding region (P38+P56)Composite
P67Low promoter+RBS+TetR Composite
P68hi promoter +RBS+lambda cI coding region (P38+P57)Composite
P69Low promoter+RBS+lambda cI repressorComposite
P70P66 and termComposite
P71P67 and termComposite
P72P68 and termComposite
P73P69 and termComposite
P74lambda cI promoter+RBS+GFP+term (P45 + P18)CompositeYes.yes
P75P74 + P1Compositeyes in both S1 and E1yes 7/22
P76P17+P45Tet QPI+GFP1778pSB1A2 (2079)3857pMB1AmpYesYesYes
P77RBS + LacI + terminatorsBBa_P03121307Failed (liquid cultures didn\'t grow)
P78RBS + LacI + terminatorsBBa_P01121310Failed (liquid cultures didn\'t grow)
P79P90+ P48Composite
P80P90 + P49Compositeno- wrong
P83P45+mtrA (w/ Pst1)Composite
P84heat sensitive Lac- YFPJ069122314pSB1A2 (2079)pMB1AmpYes, in E1.Yes, 7/9
P85heat sensitive Lac-YFPJ069112314pSB1A2 (2079)pMB1AmpYes, in E1.yes
P86pPL-PCB from UT Austinn/aP15aAmpYes in S1
P87pPL-cph1-envZ-pprotetn/aColE1CmNo
P88light-sensing EnvZBBa_I150102238pSB2K3 (4425)F\', P1 lyticKanfailed
P89RBS + LacI + terminators + pLacBBa_Q011211372pSB1A2 (2079)3451pMB1Kanfailed
P90CDF in P1 vectorpSB3K3 (2750)p15AKan
P91P89+P45: lac QPI+complete GFP4327Amp
P92P84 + P1CompositeQ04510
P93P85+P1Composite
P94P40 + mtrBComposite>4000pSB1A2 (2079)Amp
P95RBS + cph8BBa_S03419Yes
P96RBS + cph8 + terminatorsBBa_S03417Yes
P97Strong RBSBBa_B003412pSB1A2 (2079)2.1 kbpMB1AmpYesYes
P99cell deathBBa_P1010Part from MIT aligned to ref
P101pSB4C5 (with death gene)pSB4C5/I520021090pSB4C5 (3221)4311pSC101CmYesYesYesPart from MIT aligned to ref
P102P108+P45 (lac induced GFP) in p15a vector2281pSB3K3 (2750)5031P15AKanYesYesYesYes, along with 108+45#41
P103P108+P45 (lac induced GFP) in pSC101 vector2281pSB4C5 (3221)5502pSC101CmYesYesYesP103 # 41, 45
P104P75 + P63Yes 1 colonyyes 7/28
P105P87 w/o PstI site (mutated Cph)223520224257ColE1CmyesE2P105A, E5P105D
p106pER21 - plasmid for genetic recombinationn/a : from DadGentamycinYesYes
p107pKOV - plasmid for genetic recombinationn/a: from George ChurchCmYesYes
P108P3 + (P39+51) beta (lac QPI)composite1405pSB3K3 (2750)4155p15AKanyesyesyesVerified
P109RBS+full lacZI7320173093pSB1A2 (2079)5172pMB1Ampyesyesyescorrect from MIT
P110RBS+lacZalpha+termE0435534pSB4K5 (3419)3953pSC101Kanyesyesyes\"correct from MIT
\"
P111RBS+lacZalpha+termE0435534pSB4C5 (3221)3755pSC101Cmyesyesyescorrect from MIT
P112RBS+lacZalpha+termE0435534pSB1AK3 ( 3189)3723pMB1A/Kyesyesyescorrect from MIT
P113RBS+lacZalpha+termE0435534pSB4A3 (3339)3873repA pSC101Ampyesyesyescorrect from MIT
P114pJBC2 (inducible IPTG system)Yes
P115low promoter+RBS+E. coli ompR+term (from alt method)960pSB3K3 (2750)3710P15AKanYesYesompR+P1 #64, 73 correct
P116P38+P17 in pSB3K3937pSB3K3 (2750)3687P15AKanYesYesP38+17 in P1/3 #2 , 3, 5
P117P39+P17 in pSB3K3937pSB3K3 (2750)3687P15AKanYesYesP39+17 in P1/3 #1
P118low promoter+RBS+HO-pcyA+term in pSB3K31555pSB3K3 (2750)4305P15AKanYesYesHO+P1 #3
P119P63+P18Composite
P120P63 + P18 + cI (RBS primer set)Compositeverified (8/13)
P121P104 + cI (RBS primer set)verified (8/13)
P122P120 + P38
P123P120 + P39
P124P121 + P38
P125P121 + P39
P126P115 (low ompR) in pSB4C5960pSB4C5 (3221)4181pSC101CmYesYesYesYes, along with 101+115 numbers 4, 14, 25
P127P118 (low HO-pcyA) in pSB4C51555pSB4C5 (3221)4776pSC101CmYesYesYesYes, along with 101+118 numbers 61, 77
P128P97+mtrB2106pSB1A2 (2079)4185pMB1AmpYesYesYes128# 1, 5, 9, appear to have a silent SNP
P129mtrB+P632189pSB1AK3 (3189)5378Amp/KanYesYesYes129# 21, 28 appear to have a silent SNP
P130P97+mtrB+P63Composite2201pSB1AK3 (3189)5390Amp/KanYes
P131P130 in pSB4C52201pSB4C5 (3221)5422pSC101CmA and B have F>S
P132P108 in pSB4C51405pSB4C5 (3221)4626pSC101CmA, D correct
P133P116 in pSB4C5937pSB4C5 (3221)4158pSC101CmA, B correct
P134P117 in pSB4C5937pSB4C5 (3221)4158pSC101CmA, B correct
P135P39+P1302236pSB1A2 (2079)4315pMB1Amp#31, 39, 60, 70, 80 F>S, but need more checking for SNPs in mtrB
P136P38+P1302236pSB1A2 (2079)4315pMB1Amp#20 F>S, but needs checking for SNPs in mtrB
P137P135 in pSB4C52236pSB4C5 (3221)5457pSC101Cm#32- HAS MUTATION IN MIDDLE, F>S
P138P136 in pSB4C5#1, 2> check for mutations in mtrB, are they not on the same vector?
P139P130 in P133 (high constitutive promoter+tet QPI+RBS+mtrB+terminator)no
P140P108 in P131 (low constitutive promoter+lac QPI+RBS+mtrB+terminator3637 (definite)pSB4C5 (3221)6858pSC101CmYES, but there is a F>S change in mtrB
P141P38+51 (high promoter+lac QPI)1405pSB1A2 (2079)3484pMB1Amp#3, 4 correct
P142P141 in P131 (high promoter+lac QPI+RBS+mtrB+terminator)3637pSB4C5 (3221)6858pSC101CmYES but needs scrutiny (can\'t check that hi promoter is there, and SNPs in mtrB)
P143P130 in P134 (low promoter+tet QPI+RBS+mtrB+term)no
P144high mtrB in 15a38+130+12236pSB3K3 (2750)4986p15AKanyesYesYes
P145low mtrB in 15a39+130+12236pSB3K3 (2750)4986p15AKanyesYesYes
P146high lacQPI+ mtrB in 15a38+51+130+13637pSB3K3 (2750)6387p15AKanyesYesYes
P147low lacQPI+ mtrB in 15a39+51+130+13637pSB3K3 (2750)6387p15AKanyesYesYes



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