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Plasmid Files

pRFP-C-RS

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

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pRFP-C-RS Sequence and MappRFP-C-RS.dna
Map and Sequence File   
Sequence Author:  OriGene
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 EcoRI (1) AarI (6100) SgrDI (5253) HpaI (5229) CMV enhancer SnaBI (4698) BglII (4390) BclI * (4368) PshAI (3755) AsiSI - PvuI (3673) PmlI (40) BamHI (334) HindIII (380) AvrII (644) BsiWI (729) PciI (2255) DrdI (2363) AgeI (3017) BstXI (3136) BlpI (3236) AleI (3509) FseI (3603) PmeI (3609) PaeR7I - PspXI - XhoI (3614) MluI (3632) BstBI (3660) pRFP-C-RS 7454 bp
EcoRI  (1)
1 site
G A A T T C C T T A A G
AarI  (6100)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
SgrDI  (5253)
1 site
C G T C G A C G G C A G C T G C
HpaI  (5229)
1 site
G T T A A C C A A T T G
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
BclI  (4368)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
PshAI  (3755)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
AsiSI  (3673)
1 site
G C G A T C G C C G C T A G C G
PvuI  (3673)
1 site
C G A T C G G C T A G C
PmlI  (40)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
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
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.
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.
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.
BlpI  (3236)
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.
AleI  (3509)
1 site
C A C N N N N G T G G T G N N N N C A C
FseI  (3603)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°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
TurboRFP
3688 .. 4383  =  696 bp
231 amino acids  =  26.1 kDa
Product: red fluorescent protein from Entacmaea
quadricolor

mammalian codon-optimized
TurboRFP
3688 .. 4383  =  696 bp
231 amino acids  =  26.1 kDa
Product: red fluorescent protein from Entacmaea
quadricolor

mammalian codon-optimized
CmR
5266 .. 5925  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
5266 .. 5925  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
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
6100 .. 6569  =  470 bp
long terminal repeat from Moloney murine leukemia
virus
5' LTR
6100 .. 6569  =  470 bp
long terminal repeat from Moloney murine leukemia
virus
gag (truncated)
7026 .. 7442  =  417 bp
truncated MMLV gag gene lacking the start codon
gag (truncated)
7026 .. 7442  =  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
6634 .. 6833  =  200 bp
packaging signal of Moloney murine leukemia virus
(MMLV)
MMLV Ψ
6634 .. 6833  =  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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