Standard cloning vector (phagemid excised from lambda ZAP).The f1 (+) orientation allows rescue of sense strand ssDNA.pBluescript SK(+) has a reversed MCS.
Sequence Author: Stratagene
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I.M.A.G.E. Consortium Plasmids
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HomePlasmidsI.M.A.G.E. Consortium PlasmidspBluescript KS(+)
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| XmnI(2645) 1 site |
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| BsaHI is typically used at 37°C, but is even more active at 60°C. |
| ScaI(2526) 1 site |
| TatI(2524) 1 site |
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| Efficient cleavage requires at least two copies of the NmeAIII recognition sequence. Sticky ends from different NmeAIII sites may not be compatible.For full activity, add fresh S-adenosylmethionine (SAM). |
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| Efficient cleavage requires at least two copies of the BpmI recognition sequence. Sticky ends from different BpmI sites may not be compatible.After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.BpmI quickly loses activity at 37°C. |
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| Sticky ends from different BsaI sites may not be compatible.BsaI can be used between 37°C and 50°C. |
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| The 1-base overhangs produced by AhdI may be hard to ligate. Sticky ends from different AhdI sites may not be compatible. |
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| Sticky ends from different AlwNI sites may not be compatible. |
| PsiI(102) 1 site |
| BsaAI(230) 1 site |
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| Sticky ends from different DraIII sites may not be compatible. |
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| Sticky ends from different BtgZI sites may not be compatible.After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.BtgZI is typically used at 60°C, but is 75% active at 37°C. |
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| Efficient cleavage requires at least two copies of the NgoMIV recognition sequence. |
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| Efficient cleavage requires at least two copies of the NaeI recognition sequence. |
| Eco53kI(655) 1 site |
| SacI(657) 1 site |
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| Sticky ends from different BtgI sites may not be compatible. |
| AleI(663) 1 site |
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| Efficient cleavage requires at least two copies of the SacII recognition sequence. |
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| Sticky ends from different BstXI sites may not be compatible. |
| EagI(670) 1 site |
| NotI(670) 1 site |
| XbaI(677) 1 site |
| SpeI(683) 1 site |
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| After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility. |
| TspMI(695) 1 site |
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| Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present. |
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| SmaI can be used at 37°C for brief incubations. |
| PstI(705) 1 site |
| EcoRI(707) 1 site |
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| EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage. |
| HindIII(719) 1 site |
| BspDI(726) 1 site |
| ClaI(726) 1 site |
| SalI(734) 1 site |
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| Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.Sticky ends from different AccI sites may not be compatible. |
| HincII(736) 1 site |
| AbsI(740) 1 site |
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| PaeR7I does not recognize the sequence CTCTCGAG. |
| PspXI(740) 1 site |
| XhoI(740) 1 site |
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| Sticky ends from different EcoO109I sites may not be compatible. |
| PspOMI(749) 1 site |
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| ApaI can be used between 25°C and 37°C. |
| Acc65I(755) 1 site |
| KpnI(759) 1 site |
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| Sticky ends from different BspQI sites may not be compatible. |
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| Sticky ends from different SapI sites may not be compatible.SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot. |
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| Sticky ends from different AflIII sites may not be compatible. |
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| PciI is inhibited by nonionic detergents. |
| NspI(1157) 1 site |
AmpR
1973..2833=861bp
286 amino acids=31.6 kDa
2 segments
Segment 2:
1973..2764=792bp
263 amino acids=28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1973..2833=861bp
286 amino acids=31.6 kDa
2 segments
Segment 1:signal sequence
2765..2833=69bp
23 amino acids=2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1973..2833=861bp
286 amino acids=31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1214..1802=589bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1214..1802=589bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
lacZα
244..816=573bp
190 amino acids=20.7 kDa
Product: LacZα fragment of β-galactosidase
lacZα
244..816=573bp
190 amino acids=20.7 kDa
Product: LacZα fragment of β-galactosidase
AmpR promoter
2834..2938=105bp
AmpR promoter
2834..2938=105bp
lac promoter
860..890=31bp
3 segments
Segment 3:-10
860..866=7bp
promoter for the E. coli lac operon
lac promoter
860..890=31bp
3 segments
Segment 2:
867..884=18bp
promoter for the E. coli lac operon
lac promoter
860..890=31bp
3 segments
Segment 1:-35
885..890=6bp
promoter for the E. coli lac operon
lac promoter
860..890=31bp
3 segments
promoter for the E. coli lac operon
lac operator
836..852=17bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
836..852=17bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
f1 ori
6..461=456bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6..461=456bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
MCS
653..760=108bp
pBluescript multiple cloning site
MCS
653..760=108bp
pBluescript multiple cloning site
T7 promoter
626..644=19bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
626..644=19bp
promoter for bacteriophage T7 RNA polymerase
T3 promoter
773..791=19bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
773..791=19bp
promoter for bacteriophage T3 RNA polymerase
M13 fwd
603..619=17bp
common sequencing primer, one of multiple similar variants
M13 fwd
603..619=17bp
common sequencing primer, one of multiple similar variants
M13 rev
812..828=17bp
common sequencing primer, one of multiple similar variants
M13 rev
812..828=17bp
common sequencing primer, one of multiple similar variants
ORF:2103 .. 2369=267 bp
ORF:88 amino acids=9.2 kDa
ORF:244 .. 816=573 bp
ORF:190 amino acids=20.7 kDa
ORF:1973 .. 2833=861 bp
ORF:286 amino acids=31.6 kDa
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Individual Sequences & Maps
Lafmid BA
pcDNAII
pCR2.1-TOPO (linearized)
pExpress-1
pAD-GAL4
pcDNA3.1(+)
pCR3.1
pME18S-FL3
pAMP1
pcDNA3.1(-)
pCR3.1 (linearized)
pOTB7
pAMP10
pcDNA3.1 Zeo(+)
pCR4-TOPO
pOTB7-3
pBK-CMV
pCMV SPORT
pCR4-TOPO (linearized)
pOTB7a
pBluescribe (modified)
pCMV SPORT2
pCS105
pPCR-Script Amp SK(+)
pBluescript II SK(+)
pCMV SPORT4
pCS107
pRKW2
pBluescript KS(+)
pCMV SPORT6
pCS108
pSPORT 1
pBluescript SK(+)
pCMV SPORT6.1
pCS2+
pSPORT 2
pBluescript SK(-)
pCMV SPORT6ccdB
pCS22+
pT7T3D-PacI
pBluescript-FL
pCR-Blunt II-TOPO
pCS2G
pUC19
pBluescript II KS(+)
pCR-Blunt II-TOPO (linearized)
pDNR-Dual
pYX-Asc
pBluescriptR
pCR-XL-TOPO
pDNR-LIB
pZErO-2
pcDNA3
pCR-XL-TOPO (linearized)
pDONR201
pZL1
pcDNAI
pCR2.1-TOPO
pDONR223
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