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Plasmid Files

pTXB1

Bacterial expression vector that encodes a thiol-cleavable C-terminal intein followed by a chitin-binding domain.

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pTXB1 Sequence and MappTXB1.dna
Map and Sequence File   
Sequence Author:  New England Biolabs
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 T7 terminator EcoO109I (6650) BlpI (6623) BamHI (6570) PstI (6563) MfeI (6454) StuI (6395) AgeI (6379) MreI - SgrAI (6289) SacII (6132) BtgI (6129) BsmI (6100) Mxe GyrA intein AatII (6023) ZraI (6021) BsiWI (5959) BsrGI (5948) SpeI (5793) BspQI - SapI (5775) AbsI - PaeR7I - PspXI - XhoI (5761) EcoRI (5755) MCS NotI (5748) NruI (5736) BmtI (5732) NheI (5728) NdeI (5723) RBS XbaI (5683) lac operator T7 promoter BsaBI * (5622) PmeI (5609) rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator lacI promoter BclI * (4084) BstEII (3916) ApaI (3895) PspOMI (3891) BssHII (3687) EcoRV (3652) HpaI (3596) PluTI (3463) BspEI (6700) ScaI (446) PvuI (558) FspI (704) SwaI (1099) PsiI (1196) DraIII (1324) AlwNI (1901) PciI (2310) BstZ17I (2482) PflFI - Tth111I (2507) AfeI (3196) PshAI (3257) KasI (3459) NarI * (3460) SfoI (3461) pTXB1 6706 bp
EcoO109I  (6650)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
BlpI  (6623)
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  (6570)
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.
PstI  (6563)
1 site
C T G C A G G A C G T C
MfeI  (6454)
1 site
C A A T T G G T T A A C
StuI  (6395)
1 site
A G G C C T T C C G G A
AgeI  (6379)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
MreI  (6289)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (6289)
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.
SacII  (6132)
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.
BtgI  (6129)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
BsmI  (6100)
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.
AatII  (6023)
1 site
G A C G T C C T G C A G
ZraI  (6021)
1 site
G A C G T C C T G C A G
BsiWI  (5959)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
BsrGI  (5948)
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.
SpeI  (5793)
1 site
A C T A G T T G A T C A
BspQI  (5775)
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  (5775)
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.
AbsI  (5761)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (5761)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (5761)
1 site
V C T C G A G B B G A G C T C V
XhoI  (5761)
1 site
C T C G A G G A G C T C
EcoRI  (5755)
1 site
G A A T T C C T T A A G
NotI  (5748)
1 site
G C G G C C G C C G C C G G C G
NruI  (5736)
1 site
T C G C G A A G C G C T
BmtI  (5732)
1 site
G C T A G C C G A T C G
NheI  (5728)
1 site
G C T A G C C G A T C G
NdeI  (5723)
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.
XbaI  (5683)
1 site
T C T A G A A G A T C T
BsaBI  (5622)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
PmeI  (5609)
1 site
G T T T A A A C C A A A T T T G
BclI  (4084)
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  (3916)
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  (3895)
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  (3891)
1 site
G G G C C C C C C G G G
BssHII  (3687)
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.
EcoRV  (3652)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to
delete a base after cleavage.
HpaI  (3596)
1 site
G T T A A C C A A T T G
PluTI  (3463)
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.
BspEI  (6700)
1 site
T C C G G A A G G C C T
ScaI  (446)
1 site
A G T A C T T C A T G A
PvuI  (558)
1 site
C G A T C G G C T A G C
FspI  (704)
1 site
T G C G C A A C G C G T
SwaI  (1099)
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.
PsiI  (1196)
1 site
T T A T A A A A T A T T
DraIII  (1324)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
AlwNI  (1901)
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.
PciI  (2310)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (2482)
1 site
G T A T A C C A T A T G
PflFI  (2507)
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  (2507)
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.
AfeI  (3196)
1 site
A G C G C T T C G C G A
PshAI  (3257)
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.
KasI  (3459)
1 site
G G C G C C C C G C G G
NarI  (3460)
1 site
G G C G C C C C G C G G
* Blocked by Dcm methylation.
Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (3461)
1 site
G G C G C C C C G C G G
lacI
3371 .. 4453  =  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
3371 .. 4453  =  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
140 .. 1000  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   140 .. 208  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
140 .. 1000  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   209 .. 1000  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
140 .. 1000  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
Mxe GyrA intein
5776 .. 6369  =  594 bp
198 amino acids  =  21.3 kDa
Product: modified GyrA intein (Southworth et al.,
1999)
N-terminal cleavage of this intein can be induced by
thiols.
Mxe GyrA intein
5776 .. 6369  =  594 bp
198 amino acids  =  21.3 kDa
Product: modified GyrA intein (Southworth et al.,
1999)
N-terminal cleavage of this intein can be induced by
thiols.
ori
1666 .. 2254  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1666 .. 2254  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
M13 ori
1042 .. 1555  =  514 bp
M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
M13 ori
1042 .. 1555  =  514 bp
M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
rop
2623 .. 2814  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
rop
2623 .. 2814  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
CBD
6400 .. 6555  =  156 bp
52 amino acids  =  5.7 kDa
Product: chitin binding domain from chitinase A1
(Watanabe et al., 1994)
CBD
6400 .. 6555  =  156 bp
52 amino acids  =  5.7 kDa
Product: chitin binding domain from chitinase A1
(Watanabe et al., 1994)
AmpR promoter
35 .. 139  =  105 bp
AmpR promoter
35 .. 139  =  105 bp
rrnB T1 terminator
4684 .. 4770  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
4684 .. 4770  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
4867 .. 4953  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
4867 .. 4953  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
5050 .. 5136  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
5050 .. 5136  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
5233 .. 5319  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
5233 .. 5319  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
5416 .. 5502  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
5416 .. 5502  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
lacI promoter
4454 .. 4531  =  78 bp
lacI promoter
4454 .. 4531  =  78 bp
MCS
5722 .. 5775  =  54 bp
multiple cloning site
MCS
5722 .. 5775  =  54 bp
multiple cloning site
T7 terminator
6634 .. 6681  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
6634 .. 6681  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
lac operator
5656 .. 5680  =  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
5656 .. 5680  =  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).
RBS
5695 .. 5717  =  23 bp
efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)
RBS
5695 .. 5717  =  23 bp
efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)
T7 promoter
5637 .. 5655  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
5637 .. 5655  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ATG
5725 .. 5727  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
5725 .. 5727  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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