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Plasmid Files

pGFP-V-RS

HuSH-29 retroviral vector with kanamycin and puromycin resistance markers and a TurboGFP gene, for expressing an shRNA from the U6 promoter.

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pGFP-V-RS Sequence and MappGFP-V-RS.dna
Map and Sequence File   
Sequence Author:  OriGene
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 EcoRI (1) Bsu36I (6961) NruI (6015) HpaI (5229) MfeI (5221) BsrDI (5128) CMV enhancer SnaBI (4698) BglII (4390) XmnI (4145) BsaBI * (3919) ApaI (3798) DraIII (40) BamHI (334) HindIII (380) SfiI (597) AvrII (644) BsiWI (729) BspEI * (786) ScaI (1295) BspDI - ClaI (1307) PciI (2255) DrdI (2363) BssS α I (2428) ApaLI (2569) AgeI (3017) BstXI (3136) AleI (3509) PmeI (3609) PaeR7I - PspXI - XhoI (3614) MluI (3632) BstBI (3660) AclI (3664) AsiSI (3673) PspOMI (3794) pGFP-V-RS 7586 bp
EcoRI  (1)
1 site
G A A T T C C T T A A G
Bsu36I  (6961)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
NruI  (6015)
1 site
T C G C G A A G C G C T
HpaI  (5229)
1 site
G T T A A C C A A T T G
MfeI  (5221)
1 site
C A A T T G G T T A A C
BsrDI  (5128)
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.
SnaBI  (4698)
1 site
T A C G T A A T G C A T
BglII  (4390)
1 site
A G A T C T T C T A G A
XmnI  (4145)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaBI  (3919)
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.
ApaI  (3798)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
DraIII  (40)
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.
BamHI  (334)
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.
HindIII  (380)
1 site
A A G C T T T T C G A A
SfiI  (597)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
AvrII  (644)
1 site
C C T A G G G G A T C C
BsiWI  (729)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
BspEI  (786)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
ScaI  (1295)
1 site
A G T A C T T C A T G A
BspDI  (1307)
1 site
A T C G A T T A G C T A
ClaI  (1307)
1 site
A T C G A T T A G C T A
PciI  (2255)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (2363)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
BssSαI  (2428)
1 site
C A C G A G G T G C T C
ApaLI  (2569)
1 site
G T G C A C C A C G T G
AgeI  (3017)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
BstXI  (3136)
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.
AleI  (3509)
1 site
C A C N N N N G T G G T G N N N N C A C
PmeI  (3609)
1 site
G T T T A A A C C A A A T T T G
PaeR7I  (3614)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3614)
1 site
V C T C G A G B B G A G C T C V
XhoI  (3614)
1 site
C T C G A G G A G C T C
MluI  (3632)
1 site
A C G C G T T G C G C A
BstBI  (3660)
1 site
T T C G A A A A G C T T
AclI  (3664)
1 site
A A C G T T T T G C A A
AsiSI  (3673)
1 site
G C G A T C G C C G C T A G C G
PspOMI  (3794)
1 site
G G G C C C C C C G G G
KanR
5289 .. 6098  =  810 bp
269 amino acids  =  30.8 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
KanR
5289 .. 6098  =  810 bp
269 amino acids  =  30.8 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
TurboGFP
3688 .. 4383  =  696 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from
Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
TurboGFP
3688 .. 4383  =  696 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from
Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
PuroR
673 .. 1272  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
673 .. 1272  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
3' LTR
1351 .. 1944  =  594 bp
long terminal repeat from Moloney murine leukemia
virus
3' LTR
1351 .. 1944  =  594 bp
long terminal repeat from Moloney murine leukemia
virus
ori
2316 .. 2904  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2316 .. 2904  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
hGH poly(A) signal
3091 .. 3567  =  477 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
3091 .. 3567  =  477 bp
human growth hormone polyadenylation signal
5' LTR
6232 .. 6701  =  470 bp
long terminal repeat from Moloney murine leukemia
virus
5' LTR
6232 .. 6701  =  470 bp
long terminal repeat from Moloney murine leukemia
virus
gag (truncated)
7158 .. 7574  =  417 bp
truncated MMLV gag gene lacking the start codon
gag (truncated)
7158 .. 7574  =  417 bp
truncated MMLV gag gene lacking the start codon
CMV enhancer
4675 .. 5054  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
4675 .. 5054  =  380 bp
human cytomegalovirus immediate early enhancer
U6 promoter
90 .. 330  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
90 .. 330  =  241 bp
RNA polymerase III promoter for human U6 snRNA
CMV promoter
4471 .. 4674  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
4471 .. 4674  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
MMLV Ψ
6766 .. 6965  =  200 bp
packaging signal of Moloney murine leukemia virus
(MMLV)
MMLV Ψ
6766 .. 6965  =  200 bp
packaging signal of Moloney murine leukemia virus
(MMLV)
SV40 promoter
463 .. 659  =  197 bp
SV40 early promoter
SV40 promoter
463 .. 659  =  197 bp
SV40 early promoter
shRNA site
334 .. 385  =  52 bp
shRNA insertion site
shRNA site
334 .. 385  =  52 bp
shRNA insertion site
T7 promoter
4427 .. 4445  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
4427 .. 4445  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
510 .. 645  =  136 bp
SV40 origin of replication
SV40 ori
510 .. 645  =  136 bp
SV40 origin of replication
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