pInvRecA

Plasmid for transiently converting any E. coli strain to a RecA+ phenotype for phage P1-mediated transduction.

Sequence Author: Lucigen

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BaeGI - Bme1580I * (7102) ApaLI (7098) ScaI (6911) PvuI (6801) FspI (6653) NmeAIII (6579) BglI (6551) BsaI (6492) AhdI (6431) PluTI (5991) SfoI (5989) NarI (5988) KasI (5987) AflII (5824) AflIII - PciI (5066) BstXI (4416) AatII (4289) ZraI (4287) rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator HpaI (12) BsgI (131) AleI (153) AfeI (287) EcoNI (482) Bpu10I (527) Bsu36I (814) SwaI (1388) MscI * (1794) BmgBI (2134) NheI (2198) BmtI (2202) AvaI - BsoBI - PaeR7I - XhoI (2222) BlpI (2384) PmeI (2604) BsmBI - Esp3I - PshAI (2695) DrdI (2950) SgrAI (3004) PstI (3140) RsrII (3170) BglII (3177) BamHI (3412) FRT rrnB T1 terminator pInvRecA 7354 bp
BaeGI  (7102)
1 site
G K G C M C C M C G K G

Sticky ends from different BaeGI sites may not be compatible.
Bme1580I  (7102)
1 site
G K G C M C C M C G K G
* Blocked by EcoKI methylation.
Sticky ends from different Bme1580I sites may not be compatible.
ApaLI  (7098)
1 site
G T G C A C C A C G T G
ScaI  (6911)
1 site
A G T A C T T C A T G A
PvuI  (6801)
1 site
C G A T C G G C T A G C
FspI  (6653)
1 site
T G C G C A A C G C G T
NmeAIII  (6579)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

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).
BglI  (6551)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
BsaI  (6492)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
AhdI  (6431)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
PluTI  (5991)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
SfoI  (5989)
1 site
G G C G C C C C G C G G
NarI  (5988)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (5987)
1 site
G G C G C C C C G C G G
AflII  (5824)
1 site
C T T A A G G A A T T C
AflIII  (5066)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (5066)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstXI  (4416)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
AatII  (4289)
1 site
G A C G T C C T G C A G
ZraI  (4287)
1 site
G A C G T C C T G C A G
HpaI  (12)
1 site
G T T A A C C A A T T G
BsgI  (131)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
AleI  (153)
1 site
C A C N N N N G T G G T G N N N N C A C
AfeI  (287)
1 site
A G C G C T T C G C G A
EcoNI  (482)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
Bpu10I  (527)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
Bsu36I  (814)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
SwaI  (1388)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
MscI  (1794)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
BmgBI  (2134)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
NheI  (2198)
1 site
G C T A G C C G A T C G
BmtI  (2202)
1 site
G C T A G C C G A T C G
AvaI  (2222)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (2222)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
PaeR7I  (2222)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2222)
1 site
C T C G A G G A G C T C
BlpI  (2384)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
PmeI  (2604)
1 site
G T T T A A A C C A A A T T T G
BsmBI  (2695)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (2695)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
PshAI  (2695)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
DrdI  (2950)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
SgrAI  (3004)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
PstI  (3140)
1 site
C T G C A G G A C G T C
RsrII  (3170)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BglII  (3177)
1 site
A G A T C T T C T A G A
BamHI  (3412)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
FLP
783 .. 2051  =  1269 bp
422 amino acids  =  48.5 kDa
Product: site-specific recombinase
FLP is a site-specific recombinase from Saccharomyces cerevisiae. Recombination occurs at FRT sequences.
FLP
783 .. 2051  =  1269 bp
422 amino acids  =  48.5 kDa
Product: site-specific recombinase
FLP is a site-specific recombinase from Saccharomyces cerevisiae. Recombination occurs at FRT sequences.
RecA
2311 .. 3372  =  1062 bp
353 amino acids  =  38.0 kDa
Product: DNA repair protein from E. coli
RecA
2311 .. 3372  =  1062 bp
353 amino acids  =  38.0 kDa
Product: DNA repair protein from E. coli
Rep101(Ts)
4516 .. 5466  =  951 bp
316 amino acids  =  37.4 kDa
Product: temperature-sensitive version of a protein needed for replication with the pSC101 origin
Rep101(Ts)
4516 .. 5466  =  951 bp
316 amino acids  =  37.4 kDa
Product: temperature-sensitive version of a protein needed for replication with the pSC101 origin
AmpR
6358 .. 7218  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6358 .. 7149  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6358 .. 7218  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7150 .. 7218  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6358 .. 7218  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
TetR
69 .. 692  =  624 bp
207 amino acids  =  23.4 kDa
Product: tetracycline repressor TetR
TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline.
TetR
69 .. 692  =  624 bp
207 amino acids  =  23.4 kDa
Product: tetracycline repressor TetR
TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline.
pSC101 ori
5514 .. 5736  =  223 bp
low-copy replication origin that requires the Rep101 protein
pSC101 ori
5514 .. 5736  =  223 bp
low-copy replication origin that requires the Rep101 protein
AmpR promoter
7219 .. 7323  =  105 bp
AmpR promoter
7219 .. 7323  =  105 bp
rrnB T1 terminator
3562 .. 3648  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
3562 .. 3648  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
3743 .. 3829  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
3743 .. 3829  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
3924 .. 4010  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
3924 .. 4010  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
4105 .. 4191  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
4105 .. 4191  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
tetR/tetA promoters
711 .. 766  =  56 bp
overlapping promoters for bacterial tetR and tetA
tetR/tetA promoters
711 .. 766  =  56 bp
overlapping promoters for bacterial tetR and tetA
FRT
2251 .. 2284  =  34 bp
FLP-mediated recombination occurs in the 8-bp core sequence (TTTCTAGA).
FRT
2251 .. 2284  =  34 bp
FLP-mediated recombination occurs in the 8-bp core sequence (TTTCTAGA).
FRT
3425 .. 3458  =  34 bp
FLP-mediated recombination occurs in the 8-bp core sequence (TTTCTAGA).
FRT
3425 .. 3458  =  34 bp
FLP-mediated recombination occurs in the 8-bp core sequence (TTTCTAGA).
tet operator
717 .. 735  =  19 bp
bacterial operator O1 for the tetR and tetA genes
tet operator
717 .. 735  =  19 bp
bacterial operator O1 for the tetR and tetA genes
tet operator
747 .. 765  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
747 .. 765  =  19 bp
bacterial operator O2 for the tetR and tetA genes
ORF:  6488 .. 6754  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  774 .. 2051  =  1278 bp
ORF:  425 amino acids  =  48.8 kDa
ORF:  1115 .. 1468  =  354 bp
ORF:  117 amino acids  =  13.5 kDa
ORF:  2311 .. 3372  =  1062 bp
ORF:  353 amino acids  =  38.0 kDa
ORF:  4036 .. 4266  =  231 bp
ORF:  76 amino acids  =  8.8 kDa
ORF:  4516 .. 5466  =  951 bp
ORF:  316 amino acids  =  37.4 kDa
ORF:  6358 .. 7218  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  69 .. 695  =  627 bp
ORF:  208 amino acids  =  23.5 kDa
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