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Plasmid Files

pInvRecA

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

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pInvRecA Sequence and MappInvRecA.dna
Map and Sequence File   
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) BmeT110I (2223) BlpI (2384) PmeI (2604) BsmBI - 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

The sticky ends produced by AflII are hard to ligate.
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

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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
BmeT110I  (2223)
1 site
C Y C G R G G R G C Y 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.
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
   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
   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
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