Resources
Plasmid Files

pAcGHLT A

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

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

pAcGHLT A.dna
Map and Sequence File:    Download    Open   
Sequence Author:  BD Biosciences
Download Free Trial Get SnapGene Viewer


PspFI (8097) BseYI (8093) AlwNI (7988) BpmI (7440) NmeAIII (7362) PfoI (6470) DraIII (6113) FspAI (4733) SgrDI (4580) SphI (234) BstXI (1255) BbvCI - Bpu10I (1504) EcoRV (2099) EcoNI (2216) BamHI (2862) NdeI (2959) EcoRI (2974) StuI (2982) BtgI - NcoI - StyI (2986) Eco53kI (2994) SacI (2996) NotI (2999) PstI - SbfI (3010) Acc65I (3012) KpnI (3016) TspMI - XmaI (3018) SmaI (3020) BglII (3024) SnaBI (3213) AgeI (4061) SgrAI (4301) pAcGHLT A 8757 bp
PspFI  (8097)
1 site
C C C A G C G G G T C G
BseYI  (8093)
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  (7988)
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  (7440)
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  (7362)
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  (6470)
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  (6113)
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  (4733)
1 site
R T G C G C A Y Y A C G C G T R
SgrDI  (4580)
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

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
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.
NdeI  (2959)
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.
EcoRI  (2974)
1 site
G A A T T C C T T A A G
StuI  (2982)
1 site
A G G C C T T C C G G A
BtgI  (2986)
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  (2986)
1 site
C C A T G G G G T A C C
StyI  (2986)
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  (2994)
1 site
G A G C T C C T C G A G
SacI  (2996)
1 site
G A G C T C C T C G A G
NotI  (2999)
1 site
G C G G C C G C C G C C G G C G
PstI  (3010)
1 site
C T G C A G G A C G T C
SbfI  (3010)
1 site
C C T G C A G G G G A C G T C C
Acc65I  (3012)
1 site
G G T A C C C C A T G G
KpnI  (3016)
1 site
G G T A C C C C A T G G
TspMI  (3018)
1 site
C C C G G G G G G C C C
XmaI  (3018)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (3020)
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  (3024)
1 site
A G A T C T T C T A G A
SnaBI  (3213)
1 site
T A C G T A A T