pTYB21

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

Sequence Author: New England Biolabs

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HomePlasmidsBasic Cloning VectorspTYB21
SpeI (7372) PstI - SbfI (7359) EcoRI (7348) BamHI (7342) 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) Sce VMA intein 3' region MscI (6752) MfeI (6646) BmtI (6593) NheI (6589) BglII (6255) HindIII (6224) Sce VMA intein 5' region KpnI (5786) Acc65I (5782) RBS 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) BlpI (7431) T7 terminator BspEI (7508) PvuI (558) FspI (704) BsaI (861) SwaI (1099) PsiI (1196) AvaI - BsoBI (1213) 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
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
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.
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.
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.
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.
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
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)
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)
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.
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
2 segments
   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
2 segments
   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
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
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
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
ORF:  2953 .. 3588  =  636 bp
ORF:  211 amino acids  =  23.1 kDa
ORF:  4492 .. 4839  =  348 bp
ORF:  115 amino acids  =  12.9 kDa
ORF:  5725 .. 7368  =  1644 bp
ORF:  547 amino acids  =  60.7 kDa
ORF:  140 .. 1000  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  3351 .. 3614  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  4203 .. 4568  =  366 bp
ORF:  121 amino acids  =  13.1 kDa
ORF:  3371 .. 4330  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  604 .. 870  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2623 .. 2847  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  2989 .. 3315  =  327 bp
ORF:  108 amino acids  =  11.8 kDa
ORF:  4654 .. 5415  =  762 bp
ORF:  253 amino acids  =  28.4 kDa
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Individual Sequences & Maps

BlueScribe
pEASY-T1 (linearized)
pJET1.2
pSMART LCAmp
CAG promoter
pEASY-T1 Simple (linearized)
pJET1.2 blunt
pSMART LCAmp (linearized)
chicken beta-actin promoter
pEASY-T3 (linearized)
pJW168
pSMART LCKan
CLIP-tag
pEASY-T5 Zero (linearized)
pKF 18k-2
pSMART LCKan (linearized)
CMV promoter
pEX-1
pKF 19k-2
pSP64 Poly(A)
GAL1 promoter
pEX-A
pKOV
pSP72
GST
pEX-C-GST
pLATE11 (linearized)
pSP73
HaloTag
pEX-C-His
pLATE31 (linearized)
pSpark Done (linearized)
HSV TK promoter
pEX-K
pLATE51 (linearized)
pSpark I (linearized)
lac operator
pEX-N-GST
pLATE52 (linearized)
pSpark II (linearized)
lac promoter
pEX-N-His
pLEXX-AK
pSpark III (linearized)
lac UV5 promoter
pEX-N-His-GST
pLPS-B (linearized)
pSpark IV (linearized)
lacZ
pEX1
pMAL-c5E
pSpark V (linearized)
lambda
pEX2
pMAL-c5G
pSpot1
LITMUS28
pEX3
pMAL-c5X
pSpot2
LITMUS28i
pEZSeq-Amp
pMAL-c5X-His
pSpot3
LITMUS29
pEZSeq-Amp (linearized)
pMAL-c6T
pSpot4
LITMUS38
pEZSeq-Kan
pMAL-p5E
pSTBlue-1
LITMUS38i
pEZSeq-Kan (linearized)
pMAL-p5G
pSTV28
LITMUS39
pEZ BAC
pMAL-p5X
pSTV29
M13mp18
pEZ BAC (BamHI linearized)
pMCS5
pT7Blue
M13mp19
pEZ BAC (HpaI linearized)
pMiniT
pT7Blue-2
MBP
pF1A T7
pMiniT (linearized)
pT7Blue-3
pACYC177
pF1K T7
pMiniT 2.0
pT7CFE1-CFtag
pACYC184
pF25A ICE T7
pMiniT 2.0 (linearized)
pT7CFE1-CGFP-HA-His
pAN7-1
pF25K ICE T7
pMMB190
pT7CFE1-CGST-HA-His
pANT7_cGST
pF3A WG (BYDV)
pMMB206
pT7CFE1-CHA
pANT7_nHA
pF3K WG (BYDV)
pMMB207
pT7CFE1-CHis
pAUR316
pFC20A HaloTag T7 SP6
pMMB208
pT7CFE1-CMyc
pBC KS(+)
pFC20K HaloTag T7 SP6
pMMB66EH
pT7CFE1-NFtag
pBC KS(-)
pFC30A His6HaloTag T7
pMMB66HE
pT7CFE1-NHA
pBC SK(+)
pFC30K His6HaloTag T7
pMMB67EH
pT7CFE1-NHA-CHis
pBC SK(-)
pFC7A (HQ)
pMMB67HE
pT7CFE1-NHis
pBeloBAC11
pFC7K (HQ)
pNEB193
pT7CFE1-NHis-GST
pBluescript II KS(+)
pFN18A HaloTag T7
pNEB206A (linearized)
pT7CFE1-NHis-GST-CHA
pBluescript II KS(-)
pFN18K HaloTag T7
pOSIP-CH
pT7CFE1-NMyc
pBluescript II SK(+)
pFN19A HaloTag T7 SP6
pOSIP-CO
pTOP Blunt V2 (linearized)
pBluescript II SK(-)
pFN19K HaloTag T7 SP6
pOSIP-CT
pTrcHis A
pBluescript KS(+)
pFN29A His6HaloTag T7
pOSIP-KC
pTrcHis B
pBluescript KS(-)
pFN29K His6HaloTag T7
pOSIP-KH
pTrcHis C
pBluescript SK(+)
pFN2A (GST)
pOSIP-KL
pTrcHis2 A
pBluescript SK(-)
pFN2K (GST)
pOSIP-KO
pTrcHis2 B
pBR322
pFN6A (HQ)
pOSIP-KP
pTrcHis2 C
pBS KS(+)
pFN6K (HQ)
pOSIP-KT
pTV 118N
pBS KS(-)
pGEM-11Zf(+)
pOSIP-TH
pTWV228
pBS SK(+)
pGEM-11Zf(-)
pOSIP-TO
pTXB1
pBS SK(-)
pGEM-3Z
pOSIP-TT
pTYB21
pBS(+)
pGEM-3Zf(+)
pPCR-Script Amp SK(+)
pType IIs
pBS(-)
pGEM-3Zf(-)
pPCR-Script Cam SK(+)
pTZ19R
pBSK(+) Simple-Amp
pGEM-4Z
pPTR I
pTZ19U
pBSK(+) Simple-Kan
pGEM-5Zf(+)
pPTR II
pTZ57R
pcDNA3
pGEM-7Zf(+)
pRACE (linearized)
pUAST
pcDNA3.1(+)
pGEM-7Zf(-)
pRANGER-BTB-1
pUC118
pcDNA3.1(-)
pGEM-9Zf(-)
pRANGER-BTB-2
pUC119
pCold I
pGEM-T
pRANGER-BTB-3
pUC18
pCold II
pGEM-T (linearized)
pRANGER-BTB-5
pUC19
pCold III
pGEM-T Easy
pRHA-109
pUC19L
pCold IV
pGEM-T Easy (linearized)
pRHA-113
pUC57
pCold TF
pGGA
pRHA-67
pUC57-Kan
pCR-XL-2-TOPO
pGGAselect
pRSET A
pUC57-Simple
pCR-XL-2-TOPO (linearized)
PGK promoter
pRSET B
pUCmu
pDD
pGS-21a
pRSET C
pVLT31
pDNR-Dual
pH6HTC His6HaloTag T7
pSB1C3
pVLT33
pDNR-LIB
pH6HTN His6HaloTag T7
pSC101
pVLT35
pE-SUMOpro Amp
pHP45(omega)
pSEAP2-Basic
pYES1L
pE-SUMOpro Kan
pHP45(omega)-Km
pSF-Core
pZErO-2
pE-SUMOpro3 Amp
pHP45(omega)-Tc
pSMART BAC v2.0
SNAP-tag
pE-SUMOpro3 Kan
pHSG298
pSMART BAC v2.0 (BamHI linearized)
SP6 promoter
pE-SUMOstar Amp
pHSG299
pSMART BAC v2.0 (EcoRI linearized)
SV40 promoter
pE-SUMOstar Kan
pHSG396
pSMART BAC v2.0 (HindIII linearized)
T3 promoter
pEASY-Blunt (linearized)
pHSG398
pSMART FOS v2.0 (linearized)
T5 promoter
pEASY-Blunt E1 (linearized)
pHY300PLK
pSMART HCAmp
T7 promoter
pEASY-Blunt E2 (linearized)
pICOz
pSMART HCAmp (linearized)
tac promoter
pEASY-Blunt Simple (linearized)
pInvRecA
pSMART HCKan
trc promoter
pEASY-Blunt Zero (linearized)
pJAZZ-OC
pSMART HCKan (linearized)
phiX174
pEASY-Blunt3 (linearized)
pJAZZ-OK

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Insect Cell Vectors
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TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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