Resources
Plasmid Files

pTYB21

Bacterial expression vector that encodes a thiol-cleavable N-terminal intein with a chitin-binding domain.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pTYB21 Sequence and MappTYB21.dna
Map and Sequence File   
Sequence Author:  New England Biolabs
Download Free Trial Get SnapGene Viewer

 BlpI (7431) SpeI (7372) PstI - SbfI (7359) EcoRI (7348) BamHI (7342) MCS SalI - SgrDI (7336) EcoRV (7332) NotI (7323) NcoI (7316) NdeI (7310) SacI (7308) Eco53kI (7306) BspQI - SapI (7296) SacII (7188) PmlI (7177) StuI (7152) BstBI (7123) MscI (6752) CBD MfeI (6646) BmtI (6593) NheI (6589) BglII (6255) HindIII (6224) Sce VMA intein 5' region KpnI (5786) Acc65I (5782) MBP peptide XbaI (5683) lac operator T7 promoter PmeI (5609) rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator EcoNI (4566) lacI promoter MluI (4098) BstEII (3916) T7 terminator BspEI (7508) AmpR promoter PvuI (558) FspI (704) BsaI (861) SwaI (1099) PsiI (1196) AvaI - BsoBI (1213) BmeT110I (1214) DraIII (1324) PciI (2310) BstZ17I (2482) AfeI (3196) KasI (3459) NarI * (3460) SfoI (3461) PluTI (3463) BssHII (3687) PspOMI (3891) ApaI (3895) pTYB21 7514 bp
BlpI  (7431)
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.
SpeI  (7372)
1 site
A C T A G T T G A T C A
PstI  (7359)
1 site
C T G C A G G A C G T C
SbfI  (7359)
1 site
C C T G C A G G G G A C G T C C
EcoRI  (7348)
1 site
G A A T T C C T T A A G
BamHI  (7342)
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.
SalI  (7336)
1 site
G T C G A C C A G C T G
SgrDI  (7336)
1 site
C G T C G A C G G C A G C T G C
EcoRV  (7332)
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.
NotI  (7323)
1 site
G C G G C C G C C G C C G G C G
NcoI  (7316)
1 site
C C A T G G G G T A C C
NdeI  (7310)
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.
SacI  (7308)
1 site
G A G C T C C T C G A G
Eco53kI  (7306)
1 site
G A G C T C C T C G A G
BspQI  (7296)
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  (7296)
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.
SacII  (7188)
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.
PmlI  (7177)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
StuI  (7152)
1 site
A G G C C T T C C G G A
BstBI  (7123)
1 site
T T C G A A A A G C T T
MscI  (6752)
1 site
T G G C C A A C C G G T
MfeI  (6646)
1 site
C A A T T G G T T A A C
BmtI  (6593)
1 site
G C T A G C C G A T C G
NheI  (6589)
1 site
G C T A G C C G A T C G
BglII  (6255)
1 site
A G A T C T T C T A G A
HindIII  (6224)
1 site
A A G C T T T T C G A A
KpnI  (5786)
1 site
G G T A C C C C A T G G
Acc65I  (5782)
1 site
G G T A C C C C A T G G
XbaI  (5683)
1 site
T C T A G A A G A T C T
PmeI  (5609)
1 site
G T T T A A A C C A A A T T T G
EcoNI  (4566)
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  (4098)
1 site
A C G C G T T G C G C A
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.
BspEI  (7508)
1 site
T C C G G A A G G C C T
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
BsaI  (861)
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.
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
AvaI  (1213)
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  (1213)
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.
BmeT110I  (1214)
1 site
C Y C G R G G R G C Y C
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.
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
AfeI  (3196)
1 site
A G C G C T T C G C G A
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
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.
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.
PspOMI  (3891)
1 site
G G G C C C C C C G G G
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.
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
Sce VMA intein 5' region
5770 .. 6588  =  819 bp
273 amino acids  =  31.2 kDa
Product: 5' region of the intein from the yeast Vma1
subunit of the vacuolar ATPase (Chong et al., 1998)
Sce VMA intein 5' region
5770 .. 6588  =  819 bp
273 amino acids  =  31.2 kDa
Product: 5' region of the intein from the yeast Vma1
subunit of the vacuolar ATPase (Chong et al., 1998)
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
Sce VMA intein 3' region
6751 .. 7299  =  549 bp
183 amino acids  =  19.9 kDa
Product: modified 3' region of the intein from the
yeast Vma1 subunit of the vacuolar ATPase (Chong
et al., 1998)
C-terminal cleavage of this intein can be induced by
thiols.
Sce VMA intein 3' region
6751 .. 7299  =  549 bp
183 amino acids  =  19.9 kDa
Product: modified 3' region of the intein from the
yeast Vma1 subunit of the vacuolar ATPase (Chong
et al., 1998)
C-terminal cleavage of this intein can be induced by
thiols.
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
6595 .. 6747  =  153 bp
51 amino acids  =  5.6 kDa
Product: chitin binding domain from chitinase A1
(Watanabe et al., 1994)
CBD
6595 .. 6747  =  153 bp
51 amino acids  =  5.6 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
7300 .. 7377  =  78 bp
multiple cloning site
MCS
7300 .. 7377  =  78 bp
multiple cloning site
T7 terminator
7442 .. 7489  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
7442 .. 7489  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
MBP peptide
5725 .. 5754  =  30 bp
10 amino acids  =  1.2 kDa
Product: N-terminal peptide from mature
maltose-binding protein
MBP peptide
5725 .. 5754  =  30 bp
10 amino acids  =  1.2 kDa
Product: N-terminal peptide from mature
maltose-binding protein
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
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter