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Plasmid Files

pShuttle2

Vector for creating a gene expression cassette that can be transferred to a replication-deficient Ad5 genome.

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pShuttle2 Sequence and MappShuttle2.dna
Map and Sequence File   
Sequence Author:  Clontech
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 Bsu36I (17) DraIII (3627) BsrDI (3603) BspDI - ClaI (3400) SmaI (3219) TspMI - XmaI (3217) EcoNI (3180) SspI (3168) BsrFI (3135) AsiSI - PvuI (3095) BsmBI (3073) PflMI (2833) BstEII (2163) I-CeuI (20) MfeI (184) MluI (251) HincII (257) SpeI (272) NdeI (507) BmrI (562) BsaAI - SnaBI (613) NcoI - StyI (633) CMV promoter BsaI (899) NheI (918) BmtI (922) DraI - PmeI (927) PspOMI (932) EcoO109I (933) ApaI (936) XbaI (938) EagI - NotI (950) BstXI (963) Acc65I (981) KpnI (985) AflII (990) stop codons PsiI (1026) PstI (1121) EcoRI (1127) PI-SceI (1147) BspQI - SapI (1248) PciI (1364) NspI (1368) DrdI (1472) BseYI (1668) PspFI (1672) AcuI - Eco57MI (1912) pShuttle2 3974 bp
Bsu36I  (17)
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.
DraIII  (3627)
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.
BsrDI  (3603)
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.
BspDI  (3400)
1 site
A T C G A T T A G C T A
ClaI  (3400)
1 site
A T C G A T T A G C T A
SmaI  (3219)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (3217)
1 site
C C C G G G G G G C C C
XmaI  (3217)
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.
EcoNI  (3180)
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.
SspI  (3168)
1 site
A A T A T T T T A T A A
BsrFI  (3135)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI
recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its
electrophoretic mobility.
AsiSI  (3095)
1 site
G C G A T C G C C G C T A G C G
PvuI  (3095)
1 site
C G A T C G G C T A G C
BsmBI  (3073)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
PflMI  (2833)
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.
BstEII  (2163)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
I-CeuI  (20)
1 site
T A A C T A T A A C G G T C C T A A G G T A G C G A A T T G A T A T T G C C A G G A T T C C A T C G C T

I-CeuI is a homing endonuclease that can recognize a variety of
similar recognition sequences.
MfeI  (184)
1 site
C A A T T G G T T A A C
MluI  (251)
1 site
A C G C G T T G C G C A
HincII  (257)
1 site
G T Y R A C C A R Y T G
SpeI  (272)
1 site
A C T A G T T G A T C A
NdeI  (507)
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.
BmrI  (562)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the
absence of magnesium.
BsaAI  (613)
1 site
Y A C G T R R T G C A Y
SnaBI  (613)
1 site
T A C G T A A T G C A T
NcoI  (633)
1 site
C C A T G G G G T A C C
StyI  (633)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BsaI  (899)
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  (918)
1 site
G C T A G C C G A T C G
BmtI  (922)
1 site
G C T A G C C G A T C G
DraI  (927)
1 site
T T T A A A A A A T T T
PmeI  (927)
1 site
G T T T A A A C C A A A T T T G
PspOMI  (932)
1 site
G G G C C C C C C G G G
EcoO109I  (933)
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.
ApaI  (936)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
XbaI  (938)
1 site
T C T A G A A G A T C T
EagI  (950)
1 site
C G G C C G G C C G G C
NotI  (950)
1 site
G C G G C C G C C G C C G G C G
BstXI  (963)
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.
Acc65I  (981)
1 site
G G T A C C C C A T G G
KpnI  (985)
1 site
G G T A C C C C A T G G
AflII  (990)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
PsiI  (1026)
1 site
T T A T A A A A T A T T
PstI  (1121)
1 site
C T G C A G G A C G T C
EcoRI  (1127)
1 site
G A A T T C C T T A A G
PI-SceI  (1147)
1 site
A T C T A T G T C G G G T G C G G A G A A A G A G G T A A T G A A A T G G T A G A T A C A G C C C A C G C C T C T T T C T C C A T T A C T T T A C C

PI-SceI  is a homing endonuclease that can recognize a variety of
similar recognition sequences.
BspQI  (1248)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (1248)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
PciI  (1364)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NspI  (1368)
1 site
R C A T G Y Y G T A C R
DrdI  (1472)
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.
BseYI  (1668)
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.
PspFI  (1672)
1 site
C C C A G C G G G T C G
AcuI  (1912)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI
recognition sequence.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its
electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
Eco57MI  (1912)
1 site
C T G R A G ( N ) 14 N N G A C Y T C ( N ) 14

Sticky ends from different Eco57MI sites may not be compatible.
After cleavage, Eco57MI can remain bound to DNA and alter its
electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
KanR
2710 .. 3525  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
2710 .. 3525  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
ori
1425 .. 2013  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1425 .. 2013  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CMV enhancer
258 .. 637  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
258 .. 637  =  380 bp
human cytomegalovirus immediate early enhancer
CMV promoter
638 .. 841  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
638 .. 841  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 poly(A) signal
1007 .. 1088  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1007 .. 1088  =  82 bp
SV40 polyadenylation signal
MCS
918 .. 995  =  78 bp
multiple cloning site
MCS
918 .. 995  =  78 bp
multiple cloning site
T7 promoter
886 .. 904  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
886 .. 904  =  19 bp
promoter for bacteriophage T7 RNA polymerase
stop codons
996 .. 1006  =  11 bp
stop codons in all three reading frames
stop codons
996 .. 1006  =  11 bp
stop codons in all three reading frames
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