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Plasmid Files

psPAX2

Second generation lentiviral packaging vector from the Trono lab.

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psPAX2 Sequence and MappsPAX2.dna
Map and Sequence File   
Sequence Author:  Addgene
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 BsiWI (10,513) BlpI (10,481) EagI (10,337) BsaAI - SnaBI (9724) CMV enhancer PvuI (8816) FspI (8668) PspFI (7861) BseYI (7857) DrdI (7661) PsiI (7260) StuI (7219) SfiI (7173) BsaBI * (7039) PflMI (6590) BmtI (6381) NheI (6377) AleI (5593) SacI (5012) MreI - SgrAI (10,639) BspDI * - ClaI * (480) SphI (1096) BclI * (2078) SbfI (2492) BstZ17I (2578) EcoRV (2628) PflFI - Tth111I (2730) SwaI (3365) AflII (4392) XcmI (4784) Bsu36I (4964) Eco53kI (5010) psPAX2 10,668 bp
BsiWI  (10,513)
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.
BlpI  (10,481)
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.
EagI  (10,337)
1 site
C G G C C G G C C G G C
BsaAI  (9724)
1 site
Y A C G T R R T G C A Y
SnaBI  (9724)
1 site
T A C G T A A T G C A T
PvuI  (8816)
1 site
C G A T C G G C T A G C
FspI  (8668)
1 site
T G C G C A A C G C G T
PspFI  (7861)
1 site
C C C A G C G G G T C G
BseYI  (7857)
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.
DrdI  (7661)
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.
PsiI  (7260)
1 site
T T A T A A A A T A T T
StuI  (7219)
1 site
A G G C C T T C C G G A
SfiI  (7173)
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.
BsaBI  (7039)
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.
PflMI  (6590)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
BmtI  (6381)
1 site
G C T A G C C G A T C G
NheI  (6377)
1 site
G C T A G C C G A T C G
AleI  (5593)
1 site
C A C N N N N G T G G T G N N N N C A C
SacI  (5012)
1 site
G A G C T C C T C G A G
MreI  (10,639)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (10,639)
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.
BspDI  (480)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (480)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
SphI  (1096)
1 site
G C A T G C C G T A C G
BclI  (2078)
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.
SbfI  (2492)
1 site
C C T G C A G G G G A C G T C C
BstZ17I  (2578)
1 site
G T A T A C C A T A T G
EcoRV  (2628)
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.
PflFI  (2730)
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  (2730)
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.
SwaI  (3365)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
AflII  (4392)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
XcmI  (4784)
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.
Bsu36I  (4964)
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.
Eco53kI  (5010)
1 site
G A G C T C C T C G A G
HIV-1 pol
1734 .. 4745  =  3012 bp
1003 amino acids  =  113.7 kDa
Product: pol protein from human immunodeficiency
virus 1
HIV-1 pol
1734 .. 4745  =  3012 bp
1003 amino acids  =  113.7 kDa
Product: pol protein from human immunodeficiency
virus 1
AmpR
8373 .. 9233  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   8373 .. 9164  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
8373 .. 9233  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   9165 .. 9233  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
8373 .. 9233  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
7614 .. 8202  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
7614 .. 8202  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CMV enhancer
9369 .. 9748  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
9369 .. 9748  =  380 bp
human cytomegalovirus immediate early enhancer
RRE
5319 .. 5552  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
5319 .. 5552  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
SV40 promoter
7040 .. 7235  =  196 bp
SV40 early promoter
SV40 promoter
7040 .. 7235  =  196 bp
SV40 early promoter
SV40 poly(A) signal
7241 .. 7375  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
7241 .. 7375  =  135 bp
SV40 polyadenylation signal
AmpR promoter
9234 .. 9338  =  105 bp
AmpR promoter
9234 .. 9338  =  105 bp
β-globin poly(A) signal
6451 .. 6506  =  56 bp
rabbit β-globin polyadenylation signal
β-globin poly(A) signal
6451 .. 6506  =  56 bp
rabbit β-globin polyadenylation signal
HIV-1 gag
439 .. 1941  =  1503 bp
500 amino acids  =  55.9 kDa
Product: gag protein from human immunodeficiency
virus 1
HIV-1 gag
439 .. 1941  =  1503 bp
500 amino acids  =  55.9 kDa
Product: gag protein from human immunodeficiency
virus 1
SV40 ori
7086 .. 7221  =  136 bp
SV40 origin of replication
SV40 ori
7086 .. 7221  =  136 bp
SV40 origin of replication
cPPT/CTS
4430 .. 4547  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
4430 .. 4547  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
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