pAcGHLT B

Baculovirus transfer vector for expressing proteins with an N-terminal GST-6xHis-PKA-thrombin cassette. For other reading frames, use pAcGHLT A or pAcGHLT C.

Sequence Author: BD Biosciences

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HomePlasmidsInsect Cell VectorspAcGHLT B
PspFI (8095) BseYI (8091) AlwNI (7986) BpmI (7438) NmeAIII (7360) PfoI (6468) DraIII (6111) FspAI (4731) SgrDI (4578) SphI (234) BstXI (1255) BbvCI - Bpu10I (1504) baculovirus recombination region (lef2/ORF603) EcoRV (2099) EcoNI (2216) BamHI (2862) PKA site thrombin site PaeR7I - PspXI - XhoI (2966) EcoRI (2972) StuI (2980) BtgI - NcoI - StyI (2984) Eco53kI (2992) SacI (2994) NotI (2997) PstI - SbfI (3008) Acc65I (3010) KpnI (3014) TspMI - XmaI (3016) SmaI (3018) BglII (3022) SnaBI (3211) AgeI (4059) SgrAI (4299) pAcGHLT B 8755 bp
PspFI  (8095)
1 site		 C C C A G C G G G T C G
BseYI  (8091)
1 site		 C C C A G C G G G T C G
After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
AlwNI  (7986)
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.
BpmI  (7438)
1 site		 C T G G A G ( N ) 14 N N G A C C T C ( N ) 14
Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
NmeAIII  (7360)
1 site		 G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19
Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
PfoI  (6468)
1 site		 T C C N G G A A G G N C C T
Sticky ends from different PfoI sites may not be compatible.
DraIII  (6111)
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.
FspAI  (4731)
1 site		 R T G C G C A Y Y A C G C G T R
SgrDI  (4578)
1 site		 C G T C G A C G G C A G C T G C
SphI  (234)
1 site		 G C A T G C C G T A C G
BstXI  (1255)
1 site		 C C A N N N N N N T G G G G T N N N N N N A C C
Sticky ends from different BstXI sites may not be compatible.
BbvCI  (1504)
1 site		 C C T C A G C G G A G T C G
Bpu10I  (1504)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
EcoRV  (2099)
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.
EcoNI  (2216)
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.
BamHI  (2862)
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  (2966)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2966)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (2966)
1 site		 C T C G A G G A G C T C
EcoRI  (2972)
1 site		 G A A T T C C T T A A G
StuI  (2980)
1 site		 A G G C C T T C C G G A
BtgI  (2984)
1 site		 C C R Y G G G G Y R C C
Sticky ends from different BtgI sites may not be compatible.
NcoI  (2984)
1 site		 C C A T G G G G T A C C
StyI  (2984)
1 site		 C C W W G G G G W W C C
Sticky ends from different StyI sites may not be compatible.
Eco53kI  (2992)
1 site		 G A G C T C C T C G A G
SacI  (2994)
1 site		 G A G C T C C T C G A G
NotI  (2997)
1 site		 G C G G C C G C C G C C G G C G
PstI  (3008)
1 site		 C T G C A G G A C G T C
SbfI  (3008)
1 site		 C C T G C A G G G G A C G T C C
Acc65I  (3010)
1 site		 G G T A C C C C A T G G
KpnI  (3014)
1 site		 G G T A C C C C A T G G
TspMI  (3016)
1 site		 C C C G G G G G G C C C
XmaI  (3016)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (3018)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
BglII  (3022)
1 site		 A G A T C T T C T A G A
SnaBI  (3211)
1 site		 T A C G T A A T G C A T
AgeI  (4059)
1 site		 A C C G G T T G G C C A
SgrAI  (4299)
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.
baculovirus recombination region (ORF1629)
3047 .. 4505  =  1459 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
3047 .. 4505  =  1459 bp
contains part of ORF1629
AmpR
6720 .. 7580  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6720 .. 6788  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6720 .. 7580  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6789 .. 7580  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6720 .. 7580  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
baculovirus recombination region (lef2/ORF603)
1269 .. 2096  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (lef2/ORF603)
1269 .. 2096  =  828 bp
contains ORF603 and part of lef2
GST
2206 .. 2859  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
GST
2206 .. 2859  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
6xHis
2875 .. 2892  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
2875 .. 2892  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
PKA site
2899 .. 2916  =  18 bp
6 amino acids  =  658.8 Da
Product: recognition site for phosphorylation by protein kinase A
PKA site
2899 .. 2916  =  18 bp
6 amino acids  =  658.8 Da
Product: recognition site for phosphorylation by protein kinase A
thrombin site
2926 .. 2943  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
thrombin site
2926 .. 2943  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
ori
7751 .. 8339  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7751 .. 8339  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
5878 .. 6333  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
5878 .. 6333  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
AmpR promoter
6615 .. 6719  =  105 bp
AmpR promoter
6615 .. 6719  =  105 bp
polyhedrin promoter
2100 .. 2191  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
2100 .. 2191  =  92 bp
promoter for the baculovirus polyhedrin gene
MCS
2966 .. 3027  =  62 bp
multiple cloning site
MCS
2966 .. 3027  =  62 bp
multiple cloning site
ORF1629
3164 .. 4505  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
3164 .. 4505  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF603
1431 .. 2036  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF603
1431 .. 2036  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
lef2
1269 .. 1393  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
lef2
1269 .. 1393  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
ORF:  451 .. 780  =  330 bp
ORF:  109 amino acids  =  12.4 kDa
ORF:  2206 .. 3087  =  882 bp
ORF:  293 amino acids  =  33.7 kDa
ORF:  4798 .. 5388  =  591 bp
ORF:  196 amino acids  =  22.9 kDa
ORF:  5596 .. 5901  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  8737 .. 248  =  267 bp
ORF:  88 amino acids  =  10.0 kDa
ORF:  761 .. 1393  =  633 bp
ORF:  210 amino acids  =  23.9 kDa
ORF:  2805 .. 3134  =  330 bp
ORF:  109 amino acids  =  12.0 kDa
ORF:  6720 .. 7580  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1475 .. 1705  =  231 bp
ORF:  76 amino acids  =  9.3 kDa
ORF:  3164 .. 4417  =  1254 bp
ORF:  417 amino acids  =  46.0 kDa
ORF:  7184 .. 7450  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  8671 .. 422  =  507 bp
ORF:  168 amino acids  =  19.7 kDa
ORF:  1431 .. 2036  =  606 bp
ORF:  201 amino acids  =  23.6 kDa
ORF:  4461 .. 4799  =  339 bp
ORF:  112 amino acids  =  13.3 kDa
ORF:  5541 .. 5858  =  318 bp
ORF:  105 amino acids  =  12.3 kDa
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Individual Sequences & Maps

BaculoDirect C-Term Linear DNA
pBACgus-5
pIEx Bac-1
pMT BiP V5-His A
BaculoDirect N-Term Linear DNA
pBACgus-6
pIEx Bac-1 Ek_LIC (linearized)
pMT BiP V5-His B
pAB-6xHis-MBP
pBACgus4x-1
pIEx Bac-3
pMT BiP V5-His C
pAB-6xHis
pBacPAK8
pIEx Bac-3 3C_LIC (linearized)
pMT V5-His A
pAB-bee
pBacPAK9
pIEx Bac-4
pMT V5-His B
pAB-bee-8xHis
pBACsurf-1
pIEx Bac-4 Ek_LIC (linearized)
pMT V5-His C
pAB-bee-FH
pBAC-1
pIEx Bac-5
pQE-TriSystem
pAB-GST
pBAC-2cp
pIEx Bac-5 3C_LIC (linearized)
pQE-TriSystem His-Strep1
pAB-MBP
pBAC-3
pIEx-1
pQE-TriSystem His-Strep2
pAc5.1 V5-His A
pBAC-5
pIEx-10
pQE-TriSystem Strep
pAc5.1 V5-His B
pBAC-6
pIEx-10 Ek_LIC
pQE-Trisystem-5
pAc5.1 V5-His C
pBiEx-1
pIEx-10 Ek_LIC (linearized)
pQE-Trisystem-6
pAcAB3
pBiEx-2
pIEx-2
pQE-TriSystem-7
pAcAB4
pBiEx-3
pIEx-3
pTriEx-1.1
pACEBac1
pCoBlast
pIEx-4
pTriEx-1.1 Hygro
pACEBac2
pCoHygro
pIEx-5
pTriEx-1.1 Neo
pAcG2T
pDEST10
pIEx-6
pTriEx-2
pAcG3X
pDEST20
pIEx-7 Ek_LIC
pTriEx-2 Hygro
pAcGHLT A
pDEST8
pIEx-7 Ek_LIC (linearized)
pTriEx-2 Neo
pAcGHLT B
pDream2.1 MCS
pIEx-8
pTriEx-3
pAcGHLT C
pFastBac Dual
pIEx-8 Ek_LIC
pTriEx-3 Hygro
pAcGP67-A
pFastBac CT-TOPO (linearized)
pIEx-8 Ek_LIC (linearized)
pTriEx-3 Neo
pAcGP67-B
pFastBac HBM-TOPO (linearized)
pIEx-9
pTriEx-4
pAcGP67-C
pFastBac NT-TOPO (linearized)
pIEx-9 3C_LIC
pTriEx-4 Ek_LIC (linearized)
pAcHLT A
pFastBac1
pIEx-9 3C_LIC (linearized)
pTriEx-4 Hygro
pAcHLT B
pFastBacHT A
pIZ V5-His
pTriEx-4 Neo
pAcHLT C
pFastBacHT B
pIZT V5-His
pTriEx-5
pAcMP2
pFastBacHT C
pMelBac A
pTriEx-5 Ek_LIC (linearized)
pAcMP3
pI-secSUMOstar
pMelBac B
pTriEx-6
pAcSecG2T
pI-SUMOstar
pMelBac C
pTriEx-6 3C_LIC (linearized)
pAcSG2
pIB V5-His
pMIB V5-His A
pTriEx-7
pAcUW51
pIB V5-His-DEST
pMIB V5-His B
pTriEx-7 Ek_LIC (linearized)
pBAC4x-1
pIDC
pMIB V5-His C
pVL-FH
pBACgus-1
pIDK
pMT-DEST48
pVL1392
pBACgus-2cp
pIDS
pMT BioEase-DEST
pVL1393
pBACgus-3

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