Team:Harvard/Plasmids
From 2008.igem.org
(Difference between revisions)
(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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==Parts== | ==Parts== | ||
*[http://spreadsheets.google.com/ccc?key=pwvxAM8xwIq37aJ-qKhmUag&hl=en Google Spreadsheet] | *[http://spreadsheets.google.com/ccc?key=pwvxAM8xwIq37aJ-qKhmUag&hl=en Google Spreadsheet] | ||
- | {| | + | {| |
- | | align="center" style="background:# | + | | align="center" style="background:#f0f0f0;"|'''Internal name''' |
- | | align="center" style="background:# | + | | align="center" style="background:#f0f0f0;"|'''Description''' |
- | | align="center" style="background:# | + | | align="center" style="background:#f0f0f0;"|'''Registry Name''' |
- | | align="center" style="background:# | + | | align="center" style="background:#f0f0f0;"|'''Part Size''' |
- | | align="center" style="background:# | + | | align="center" style="background:#f0f0f0;"|'''Vector (size)''' |
- | | align="center" style="background:# | + | | align="center" style="background:#f0f0f0;"|'''Plasmid size (vector + part)''' |
- | | align="center" style="background:# | + | | align="center" style="background:#f0f0f0;"|'''Origin''' |
- | | align="center" style="background:# | + | | align="center" style="background:#f0f0f0;"|'''Marker''' |
- | | align="center" style="background:# | + | | align="center" style="background:#f0f0f0;"|'''Transformed?''' |
- | | align="center" style="background:# | + | | align="center" style="background:#f0f0f0;"|'''Miniprep''' |
+ | | align="center" style="background:#f0f0f0;"|'''Glycerol Stock''' | ||
+ | | align="center" style="background:#f0f0f0;"|'''Sequenced?''' | ||
|- | |- | ||
- | | P1 || | + | | 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 || | + | | P2 ||Med promoter+RBS+ GFP+term||J23150/I20269||919||pSB3K3 (2750)||3669||p15A||Kan||in TOP10, not S1||Yes||||P2E2 (tube labeled 2B) |
|- | |- | ||
- | | P3 || | + | | 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 | + | | P4 ||Low-Medium copy vector||pACYC duet||4008||||||p15a||Cm||in S1 and TOP10||Yes, for both strains.||Yes for both strains.|| |
|- | |- | ||
- | | P5 | + | | P5 ||Low-Medium copy vector (w/ death gene)||pSB3K3||2750||||||p15a||Kan||in E3||Yes||Yes (B- done 6/30. A-done 7/1)|| |
|- | |- | ||
- | | | + | | P11 ||Light responsive system, dual regulation||BBa_M30109||4333||pSB1AC3 (3055)||7388||pMB1||Amp/Cm||Failed|||||| |
|- | |- | ||
- | | | + | | P13 ||Low copy plasmid||pCDFDuet-1||3781||||||CloDF13||Sm||in S1||Yes, 435 ng/uL||Yes|| |
|- | |- | ||
- | | | + | | P15 ||pTet+RBS+GFP+term||BBa_I13522||937||BBa_J23006 (3279)||||ColE1||Amp||in E2||Yes||No||P15A, P15B |
|- | |- | ||
- | | | + | | P16 ||Tet repressible promoter||BBa_R0040||54||||||pMB1||Amp||in E2||Yes||No|| |
|- | |- | ||
- | | | + | | 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 |
|- | |- | ||
- | | | + | | 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) |
|- | |- | ||
- | | | + | | 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 |
|- | |- | ||
- | | | + | | P21 ||pACYC (P4) w/ biobricks||A1 (pACYC)||???||||||???||???||in S1 and TOP10||Yes||Yes, for both strains.|| |
|- | |- | ||
- | | | + | | P22 ||ompR controlled promoter for ompC (long)||BBa_R0082||108||||||pMB1||Amp||||||||WRONGWRONGWRONG: DID NOT ALIGN TO REF |
|- | |- | ||
- | | | + | | P23 ||ompR controlled promoter for ompC (short)||BBa_R0083||78||||||pMB1||Amp||in E2||Yes||No||WRONGWRONGWRONG: DID NOT ALIGN TO REF |
|- | |- | ||
- | | | + | | P24 ||ompR controlled promoter for ompF||BBa_R0084||108||||||pMB1||Amp||in E2||Yes||No||WRONGWRONGWRONG: DID NOT ALIGN TO REF |
|- | |- | ||
- | | | + | | P25 ||Promoter activated by OmpR-P with the reporter GFP||BBa_J58102||991||||||pMB1||Amp||||||||WRONGWRONGWRONG: DID NOT ALIGN TO REF |
|- | |- | ||
- | | | + | | 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 |
|- | |- | ||
- | | | + | | P27 ||repressor plasmid w/ constitutive expression of LacI and TetR||pZS4int2||||||||pSC101||Sm||Yes||Yes (6/26)||Yes (6/26)|| |
|- | |- | ||
- | | | + | | P28 ||GFP w/ PLtctO-1||pZE21||||||||ColE1||Kan||Yes||Yes (6/26)||Yes (6/26)|| |
|- | |- | ||
- | | | + | | P29 ||Venus (mutant YFP) PL (Plambda)||pZS*2R||||||||pSC101*||Kan||Yes||Yes (6/26)||Yes (6/26)|| |
|- | |- | ||
- | | | + | | P30 ||YFP w/ PLlacO-1||pZA32||||||||p15A||Cm||Yes||Yes (6/26)||Yes (6/26)|| |
|- | |- | ||
- | | | + | | P31 ||Venus w/ PLlacO-1||pZS*1R||||||||pSC101*||Amp||Yes||Yes (6/26)||Yes (6/26)|| |
|- | |- | ||
- | | | + | | P32 ||Venus w/ PLlacO-1||pZA32||||||||p15A||Cm||Yes||Yes (6/26)||Yes (6/26)|| |
|- | |- | ||
- | | | + | | P33||Venus w/ PLtctO-1||pZA31||||||||p15A||Cm||Yes||Yes (6/26)||Yes (6/26)|| |
|- | |- | ||
- | | | + | | P34 ||GFP w/ PLlacO-1||pZE12||||||||ColE1||Amp||Yes||Yes (6/26)||Yes (6/26)|| |
|- | |- | ||
- | | | + | | P35 ||accessory ||pLDR8||||||||||||Yes||Yes (6/26)||Yes (6/26)|| |
|- | |- | ||
- | | | + | | P36 ||same as P20||BBa_I763004||||||||||Kan/Amp|||||||| |
|- | |- | ||
- | | | + | | P37 ||Constructive expression LacI w/ pTetR promoter||BBa_I722007||||||||||Amp||Failed (liquid cultures didn\'t grow)|||||| |
|- | |- | ||
- | | | + | | 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)|| |
|- | |- | ||
- | | | + | | 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)|| |
|- | |- | ||
- | | | + | | 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)|| |
|- | |- | ||
- | | | + | | 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)|| |
|- | |- | ||
- | | | + | | P42 ||LacI coding region||\"BBa_C0012 |
|- | |- | ||
- | | | + | | \"||1128||||||||Amp||Failed (liquid cultures didn\'t grow)|||||| |
|- | |- | ||
- | | | + | | P43 ||TetR coding region||BBa_C0040||660||||||||Amp||2007 (A grew)||Yes||2007 (A-done 6/30)||P43A E1 07 |
|- | |- | ||
- | | | + | | P44 ||cI lambda||\"BBa_C0051 |
|- | |- | ||
- | | | + | | \"||750||||||||Amp||2007 (A grew)||Yes||2007 (A-done 6/30)||P44A E1 07 |
|- | |- | ||
- | | | + | | 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 |
|- | |- | ||
- | | | + | | P46 ||Base Vector||BBa_I51020||||||||||Amp||A and B grew.||Yes||A and B -done 6/30|| |
|- | |- | ||
- | | | + | | 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 |
|- | |- | ||
- | | | + | | 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 |
|- | |- | ||
- | | | + | | P51||LacI QPI w/ RBS & promoter||BBa_Q04121||1370||pSB2K3 (4425)||5795||pMB1||Kan||||||||cells from MIT correct |
|- | |- | ||
- | | | + | | P52||cI QPI w/ RBS & promoter||BBa_Q04510||987||pSB2K3 (4425)||5412||pMB1||Kan|||||||| |
|- | |- | ||
- | | | + | | 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)|| |
|- | |- | ||
- | | | + | | P54||PoPS --> LacI||BBa_P0412||1308||||||||Amp||2007 (A grew)||Yes. (7/3).||2007 (A done 7/3)|| |
|- | |- | ||
- | | | + | | 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)|| |
|- | |- | ||
- | | | + | | 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|| |
|- | |- | ||
- | | | + | | 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|| |
|- | |- | ||
- | | | + | | 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|| |
|- | |- | ||
- | | | + | | 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|| |
|- | |- | ||
- | | | + | | P60||||Composite |||||||||||||||||| |
|- | |- | ||
- | | | + | | P61||Terminator||BBa_B0011||||||||||||2008 (A grew.)||Yes for 2008 A.||2008 (A done 7/4)|| |
|- | |- | ||
- | | | + | | 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)|| |
|- | |- | ||
- | | | + | | 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|| | ||
+ | |- | ||
+ | | | ||
|} | |} |
Revision as of 02:49, 27 October 2008
Vectors
- pACYC-Duet vector map
- [http://partsregistry.org/wiki/index.php?title=Part:pSB3K3 pSB3K3]
Parts
- [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+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 | Med promoter+RBS+ GFP+term | J23150/I20269 | 919 | pSB3K3 (2750) | 3669 | p15A | Kan | in TOP10, not S1 | Yes | P2E2 (tube labeled 2B) | |
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 | Low-Medium copy vector | pACYC duet | 4008 | p15a | Cm | in S1 and TOP10 | Yes, for both strains. | Yes for both strains. | |||
P5 | Low-Medium copy vector (w/ death gene) | pSB3K3 | 2750 | p15a | Kan | in E3 | Yes | Yes (B- done 6/30. A-done 7/1) | |||
P11 | Light responsive system, dual regulation | BBa_M30109 | 4333 | pSB1AC3 (3055) | 7388 | pMB1 | Amp/Cm | Failed | |||
P13 | Low copy plasmid | pCDFDuet-1 | 3781 | CloDF13 | Sm | in S1 | Yes, 435 ng/uL | Yes | |||
P15 | pTet+RBS+GFP+term | BBa_I13522 | 937 | BBa_J23006 (3279) | ColE1 | Amp | in E2 | Yes | No | P15A, P15B | |
P16 | Tet repressible promoter | BBa_R0040 | 54 | pMB1 | Amp | in E2 | Yes | No | |||
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 |
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) | ||
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 | ||
P21 | pACYC (P4) w/ biobricks | A1 (pACYC) | ??? | ??? | ??? | in S1 and TOP10 | Yes | Yes, for both strains. | |||
P22 | ompR controlled promoter for ompC (long) | BBa_R0082 | 108 | pMB1 | Amp | WRONGWRONGWRONG: DID NOT ALIGN TO REF | |||||
P23 | ompR controlled promoter for ompC (short) | BBa_R0083 | 78 | pMB1 | Amp | in E2 | Yes | No | WRONGWRONGWRONG: DID NOT ALIGN TO REF | ||
P24 | ompR controlled promoter for ompF | BBa_R0084 | 108 | pMB1 | Amp | in E2 | Yes | No | WRONGWRONGWRONG: DID NOT ALIGN TO REF | ||
P25 | Promoter activated by OmpR-P with the reporter GFP | BBa_J58102 | 991 | pMB1 | Amp | WRONGWRONGWRONG: DID NOT ALIGN TO REF | |||||
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 | ||
P27 | repressor plasmid w/ constitutive expression of LacI and TetR | pZS4int2 | pSC101 | Sm | Yes | Yes (6/26) | Yes (6/26) | ||||
P28 | GFP w/ PLtctO-1 | pZE21 | ColE1 | Kan | Yes | Yes (6/26) | Yes (6/26) | ||||
P29 | Venus (mutant YFP) PL (Plambda) | pZS*2R | pSC101* | Kan | Yes | Yes (6/26) | Yes (6/26) | ||||
P30 | YFP w/ PLlacO-1 | pZA32 | p15A | Cm | Yes | Yes (6/26) | Yes (6/26) | ||||
P31 | Venus w/ PLlacO-1 | pZS*1R | pSC101* | Amp | Yes | Yes (6/26) | Yes (6/26) | ||||
P32 | Venus w/ PLlacO-1 | pZA32 | p15A | Cm | Yes | Yes (6/26) | Yes (6/26) | ||||
P33 | Venus w/ PLtctO-1 | pZA31 | p15A | Cm | Yes | Yes (6/26) | Yes (6/26) | ||||
P34 | GFP w/ PLlacO-1 | pZE12 | ColE1 | Amp | Yes | Yes (6/26) | Yes (6/26) | ||||
P35 | accessory | pLDR8 | Yes | Yes (6/26) | Yes (6/26) | ||||||
P36 | same as P20 | BBa_I763004 | Kan/Amp | ||||||||
P37 | Constructive expression LacI w/ pTetR promoter | BBa_I722007 | Amp | Failed (liquid cultures didn\'t grow) | |||||||
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) | ||||
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) | ||||
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) | ||||
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) | ||||||
P42 | LacI coding region | \"BBa_C0012 | |||||||||
\" | 1128 | Amp | Failed (liquid cultures didn\'t grow) | ||||||||
P43 | TetR coding region | BBa_C0040 | 660 | Amp | 2007 (A grew) | Yes | 2007 (A-done 6/30) | P43A E1 07 | |||
P44 | cI lambda | \"BBa_C0051 | |||||||||
\" | 750 | Amp | 2007 (A grew) | Yes | 2007 (A-done 6/30) | P44A E1 07 | |||||
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 |
P46 | Base Vector | BBa_I51020 | Amp | A and B grew. | Yes | A and B -done 6/30 | |||||
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 | |||
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 | |
P51 | LacI QPI w/ RBS & promoter | BBa_Q04121 | 1370 | pSB2K3 (4425) | 5795 | pMB1 | Kan | cells from MIT correct | |||
P52 | cI QPI w/ RBS & promoter | BBa_Q04510 | 987 | pSB2K3 (4425) | 5412 | pMB1 | Kan | ||||
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) | ||||
P54 | PoPS --> LacI | BBa_P0412 | 1308 | Amp | 2007 (A grew) | Yes. (7/3). | 2007 (A done 7/3) | ||||
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) | ||||
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 | ||||||
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 | |||||
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 | ||||
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 | ||||
P60 | Composite | ||||||||||
P61 | Terminator | BBa_B0011 | 2008 (A grew.) | Yes for 2008 A. | 2008 (A done 7/4) | ||||||
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) | ||||||
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 | |