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Plasmid Files

p15Tv-L

Bacterial vector with a negative selection marker, for expressing a protein with an N-terminal 6xHis-TEV cassette. Also known as p15Tv-LIC.

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p15Tv-L Sequence and Mapp15Tv-L.dna
Map and Sequence File   
Sequence Author:  Structural Genomics Consortium
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 BlpI (7476) BamHI (7423) PaeR7I - PspXI - XhoI (7418) BseRI (7394) SacII (6980) KpnI (6962) Acc65I (6958) SnaBI (6650) BsrGI (6581) AanI - PsiI (6541) StuI * (6142) BseRI - NdeI (5381) 6xHis ATG NcoI (5315) XbaI (5276) lac operator T7 promoter BglII (5210) SgrAI (5169) SphI (5021) EcoNI (4956) MluI (4488) BclI * (4474) BstEII (4306) ApaI (4285) PspOMI (4281) BssHII (4077) tet promoter EcoRI (7744) SspI (191) ScaI (515) PvuI (627) PstI (754) BsaI (930) AhdI (996) AlwNI (1475) BspQI - SapI (2001) PflFI - Tth111I (2142) FspAI (2907) BsmI (3006) NruI (3389) PshAI (3647) p15Tv-L 7746 bp
BlpI  (7476)
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.
BamHI  (7423)
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.
PaeR7I  (7418)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (7418)
1 site
V C T C G A G B B G A G C T C V
XhoI  (7418)
1 site
C T C G A G G A G C T C
BseRI  (7394)
2 sites
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
SacII  (6980)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII
recognition sequence.
KpnI  (6962)
1 site
G G T A C C C C A T G G
Acc65I  (6958)
1 site
G G T A C C C C A T G G
SnaBI  (6650)
1 site
T A C G T A A T G C A T
BsrGI  (6581)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
AanI  (6541)
1 site
T T A T A A A A T A T T
PsiI  (6541)
1 site
T T A T A A A A T A T T
StuI  (6142)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
BseRI  (5381)
2 sites
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
NdeI  (5381)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
NcoI  (5315)
1 site
C C A T G G G G T A C C
XbaI  (5276)
1 site
T C T A G A A G A T C T
BglII  (5210)
1 site
A G A T C T T C T A G A
SgrAI  (5169)
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.
SphI  (5021)
1 site
G C A T G C C G T A C G
EcoNI  (4956)
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.
MluI  (4488)
1 site
A C G C G T T G C G C A
BclI  (4474)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BstEII  (4306)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
ApaI  (4285)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (4281)
1 site
G G G C C C C C C G G G
BssHII  (4077)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
EcoRI  (7744)
1 site
G A A T T C C T T A A G
SspI  (191)
1 site
A A T A T T T T A T A A
ScaI  (515)
1 site
A G T A C T T C A T G A
PvuI  (627)
1 site
C G A T C G G C T A G C
PstI  (754)
1 site
C T G C A G G A C G T C
BsaI  (930)
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  (996)
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.
AlwNI  (1475)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
BspQI  (2001)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (2001)
1 site
G C T C T T C N C G A G A A G N N N N

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.
PflFI  (2142)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (2142)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
FspAI  (2907)
1 site
R T G C G C A Y Y A C G C G T R
BsmI  (3006)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
NruI  (3389)
1 site
T C G C G A A G C G C T
PshAI  (3647)
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.
SacB
5901 .. 7322  =  1422 bp
473 amino acids  =  53.0 kDa
   Segment 1:  signal peptide  
   5901 .. 5987  =  87 bp
   29 amino acids  =  3.0 kDa
Product: secreted levansucrase that renders
bacterial growth sensitive to sucrose
negative selection marker
SacB
5901 .. 7322  =  1422 bp
473 amino acids  =  53.0 kDa
   Segment 2:  
   5988 .. 7322  =  1335 bp
   444 amino acids  =  50.0 kDa
Product: secreted levansucrase that renders
bacterial growth sensitive to sucrose
negative selection marker
SacB
5901 .. 7322  =  1422 bp
473 amino acids  =  53.0 kDa
2 segments
Product: secreted levansucrase that renders
bacterial growth sensitive to sucrose
negative selection marker
lacI
3761 .. 4843  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
lacI
3761 .. 4843  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
AmpR
209 .. 1069  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   209 .. 277  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
209 .. 1069  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   278 .. 1069  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
209 .. 1069  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1240 .. 1828  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1240 .. 1828  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
sacB promoter
5455 .. 5900  =  446 bp
sacB promoter and control region
sacB promoter
5455 .. 5900  =  446 bp
sacB promoter and control region
rop
2258 .. 2449  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
rop
2258 .. 2449  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
AmpR promoter
104 .. 208  =  105 bp
AmpR promoter
104 .. 208  =  105 bp
lacI promoter
4844 .. 4921  =  78 bp
lacI promoter
4844 .. 4921  =  78 bp
T7 terminator
7487 .. 7534  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
7487 .. 7534  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
loxP
5407 .. 5440  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
5407 .. 5440  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
tet promoter
7709 .. 7737  =  29 bp
   Segment 3:  -10  
   7709 .. 7714  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
7709 .. 7737  =  29 bp
   Segment 2:  
   7715 .. 7731  =  17 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
7709 .. 7737  =  29 bp
   Segment 1:  -35  
   7732 .. 7737  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
7709 .. 7737  =  29 bp
3 segments
E. coli promoter for tetracycline efflux protein gene
lac operator
5249 .. 5273  =  25 bp
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
5249 .. 5273  =  25 bp
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).
TEV Site
5359 .. 5379  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV Site
5359 .. 5379  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
T7 promoter
5230 .. 5248  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
5230 .. 5248  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
5329 .. 5346  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
5329 .. 5346  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
5304 .. 5309  =  6 bp
ribosome binding site
RBS
5304 .. 5309  =  6 bp
ribosome binding site
ATG
5317 .. 5319  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
5317 .. 5319  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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