pVAX1

Nonoptimized first generation DNA vaccine vector.

Sequence Author: Thermo Fisher (Invitrogen)

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PciI (2990) ApaLI (2676) AlwNI (2581) PmlI (2211) RsrII (1870) NaeI (1856) NgoMIV (1854) BsrDI (1587) NmeAIII (1578) NruI (9) MluI (29) HincII (35) NdeI (285) SnaBI (391) BtsαI (639) BsaI (677) NheI (696) BmtI (700) PmeI (705) AflII (709) HindIII (712) Acc65I (718) KpnI (722) BamHI (730) BstXI (749) ApoI - EcoRI (753) PstI (762) EcoRV (765) BstXI (775) NotI (780) AvaI - BmeT110I - BsoBI - PaeR7I - PspXI - XhoI (786) XbaI (792) EcoO109I - PspOMI (798) ApaI (802) PmeI (807) BbsI (1018) XcmI (1170) BsaBI * (1213) MscI (1436) FspI (1456) PflFI - Tth111I (1472) pVAX1 2999 bp
PciI  (2990)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
ApaLI  (2676)
1 site
G T G C A C C A C G T G
AlwNI  (2581)
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.
PmlI  (2211)
1 site
C A C G T G G T G C A C
RsrII  (1870)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
NaeI  (1856)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV  (1854)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
BsrDI  (1587)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
NmeAIII  (1578)
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).
NruI  (9)
1 site
T C G C G A A G C G C T
MluI  (29)
1 site
A C G C G T T G C G C A
HincII  (35)
1 site
G T Y R A C C A R Y T G
NdeI  (285)
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.
SnaBI  (391)
1 site
T A C G T A A T G C A T
BtsαI  (639)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsαI sites may not be compatible.
BsaI  (677)
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.
NheI  (696)
1 site
G C T A G C C G A T C G
BmtI  (700)
1 site
G C T A G C C G A T C G
PmeI  (705)
2 sites
G T T T A A A C C A A A T T T G
AflII  (709)
1 site
C T T A A G G A A T T C
HindIII  (712)
1 site
A A G C T T T T C G A A
Acc65I  (718)
1 site
G G T A C C C C A T G G
KpnI  (722)
1 site
G G T A C C C C A T G G
BamHI  (730)
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.
BstXI  (749)
2 sites
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.
ApoI  (753)
1 site
R A A T T Y Y T T A A R

ApoI is typically used at 50°C, but is 50% active at 37°C.
EcoRI  (753)
1 site
G A A T T C C T T A A G
PstI  (762)
1 site
C T G C A G G A C G T C
EcoRV  (765)
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.
BstXI  (775)
2 sites
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.
NotI  (780)
1 site
G C G G C C G C C G C C G G C G
AvaI  (786)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BmeT110I  (786)
1 site
C Y C G R G G R G C Y C
BsoBI  (786)
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.
PaeR7I  (786)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (786)
1 site
V C T C G A G B B G A G C T C V
XhoI  (786)
1 site
C T C G A G G A G C T C
XbaI  (792)
1 site
T C T A G A A G A T C T
EcoO109I  (798)
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.
PspOMI  (798)
1 site
G G G C C C C C C G G G
ApaI  (802)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PmeI  (807)
2 sites
G T T T A A A C C A A A T T T G
BbsI  (1018)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
XcmI  (1170)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
BsaBI  (1213)
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.
MscI  (1436)
1 site
T G G C C A A C C G G T
FspI  (1456)
1 site
T G C G C A A C G C G T
PflFI  (1472)
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  (1472)
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.
NeoR/KanR
1226 .. 2020  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
1226 .. 2020  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
2346 .. 2934  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2346 .. 2934  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
36 .. 415  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
36 .. 415  =  380 bp
human cytomegalovirus immediate early enhancer
bGH poly(A) signal
829 .. 1053  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
829 .. 1053  =  225 bp
bovine growth hormone polyadenylation signal
CMV promoter
416 .. 619  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
416 .. 619  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
MCS
696 .. 811  =  116 bp
multiple cloning site
MCS
696 .. 811  =  116 bp
multiple cloning site
T7 promoter
664 .. 682  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
664 .. 682  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  1226 .. 2020  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  1398 .. 1784  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  1535 .. 2068  =  534 bp
ORF:  177 amino acids  =  19.9 kDa
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