Team:Harvard/Plasmids

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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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=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]
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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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