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.

Sequence Author: Structural Genomics Consortium

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HomePlasmidsStructural Genomics Vectorsp15Tv-L
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
2 segments
   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
2 segments
   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
2 segments
   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
2 segments
   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
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
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
TEV Site
5359 .. 5379  =  21 bp
7 amino acids  =  869.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV Site
5359 .. 5379  =  21 bp
7 amino acids  =  869.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
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
3 segments
   Segment 3:  -10  
   7709 .. 7714  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
7709 .. 7737  =  29 bp
3 segments
   Segment 2:  
   7715 .. 7731  =  17 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
7709 .. 7737  =  29 bp
3 segments
   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).
T7 promoter
5230 .. 5248  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
5230 .. 5248  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
5304 .. 5309  =  6 bp
ribosome binding site
RBS
5304 .. 5309  =  6 bp
ribosome binding site
ORF:  3283 .. 3978  =  696 bp
ORF:  231 amino acids  =  25.4 kDa
ORF:  4882 .. 5145  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  209 .. 1069  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  2481 .. 2849  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  5019 .. 5258  =  240 bp
ORF:  79 amino acids  =  8.0 kDa
ORF:  5901 .. 7322  =  1422 bp
ORF:  473 amino acids  =  53.0 kDa
ORF:  673 .. 939  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3088 .. 3705  =  618 bp
ORF:  205 amino acids  =  21.8 kDa
ORF:  3741 .. 4004  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  4593 .. 5093  =  501 bp
ORF:  166 amino acids  =  17.5 kDa
ORF:  2258 .. 2482  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  2846 .. 3184  =  339 bp
ORF:  112 amino acids  =  12.6 kDa
ORF:  3761 .. 4720  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
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Individual Sequences & Maps

p11
pMCSG19B (linearized)
pMCSG41
pMCSG68 (linearized)
p15Tv-L
pMCSG19C
pMCSG41 (linearized)
pMCSG69
p283
pMCSG19C (linearized)
pMCSG42
pMCSG69 (linearized)
pBEN1-SGC
pMCSG20
pMCSG42 (linearized)
pMCSG7
pBEN1-SGC (linearized)
pMCSG20 (linearized)
pMCSG43
pMCSG7 (linearized)
pDB-HisGST
pMCSG21
pMCSG43 (linearized)
pMCSG70
pDB.GST
pMCSG21 (linearized)
pMCSG46
pMCSG70 (linearized)
pDB.His.MBP
pMCSG22
pMCSG46 (linearized)
pMCSG71
pDB.His.NusA
pMCSG22 (linearized)
pMCSG48
pMCSG71 (linearized)
pDB.His.TRX
pMCSG23
pMCSG48 (linearized)
pMCSG73
pDB.TRX
pMCSG23 (linearized)
pMCSG49
pMCSG73 (linearized)
pEU-GST-FV
pMCSG24
pMCSG49 (linearized)
pMCSG75
pEU-His-FV
pMCSG24 (linearized)
pMCSG50
pMCSG75 (linearized)
pEU-HSBC
pMCSG25
pMCSG50 (linearized)
pMCSG76
pFB-LIC-Bse
pMCSG25 (linearized)
pMCSG51
pMCSG76 (linearized)
pFB-LIC-Bse (linearized)
pMCSG26
pMCSG51 (linearized)
pMCSG77
pH3c-LIC
pMCSG26 (linearized)
pMCSG52
pMCSG77 (linearized)
pH3c-LIC (linearized)
pMCSG27
pMCSG52 (linearized)
pMCSG79
pHM3C-LIC
pMCSG27 (linearized)
pMCSG53
pMCSG79 (linearized)
pHM3C-LIC (linearized)
pMCSG28
pMCSG53 (linearized)
pMCSG8
pLIC-CH
pMCSG28 (linearized)
pMCSG54
pMCSG8 (linearized)
pLIC-CH (linearized)
pMCSG29
pMCSG54 (linearized)
pMCSG81
pLIC-EGFP
pMCSG29 (linearized)
pMCSG55
pMCSG81 (linearized)
pLIC-EGFP (linearized)
pMCSG30
pMCSG55 (linearized)
pMCSG9
pLIC-SGC1
pMCSG30 (linearized)
pMCSG56
pMCSG9 (linearized)
pLIC-SGC1 (linearized)
pMCSG31
pMCSG56 (linearized)
pNH-TrxT
pMCentr1
pMCSG31 (linearized)
pMCSG57
pNH-TrxT (linearized)
pMCentr1 (linearized)
pMCSG32
pMCSG57 (linearized)
pNIC-CH
pMCentr2
pMCSG32 (linearized)
pMCSG58
pNIC-CH (linearized)
pMCentr2 (linearized)
pMCSG33
pMCSG58 (linearized)
pNIC-CTHF
pMCentr3
pMCSG33 (linearized)
pMCSG59
pNIC-CTHF (linearized)
pMCentr3 (linearized)
pMCSG34
pMCSG59 (linearized)
pNIC-Zb
pMCSG10
pMCSG34 (linearized)
pMCSG60
pNIC-Zb (linearized)
pMCSG10 (linearized)
pMCSG34B
pMCSG60 (linearized)
pNIC28-Bsa4
pMCSG11
pMCSG34B (linearized)
pMCSG61
pNIC28-Bsa4 (linearized)
pMCSG11 (linearized)
pMCSG35
pMCSG61 (linearized)
pNYCOMPS-LIC-FH10T+ (N term) - linearized
pMCSG12
pMCSG35 (linearized)
pMCSG62
pNYCOMPS-LIC-FH10T+ (N term)
pMCSG12 (linearized)
pMCSG35B
pMCSG62 (linearized)
pSpeedET
pMCSG13
pMCSG35B (linearized)
pMCSG63
pVP13
pMCSG13 (linearized)
pMCSG37
pMCSG63 (linearized)
pVP16
pMCSG14
pMCSG37 (linearized)
pMCSG64
pVP33A
pMCSG14 (linearized)
pMCSG38
pMCSG64 (linearized)
pVP33K
pMCSG17
pMCSG38 (linearized)
pMCSG65
pVP56A
pMCSG17 (linearized)
pMCSG38C
pMCSG65 (linearized)