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Plasmid Files

pAAVS1-BSD-DNR

Mammalian donor vector with a blasticidin resistance marker for transgene integration at the AAVS1 locus in the human genome. Use together with pCas‑Guide-AAVS1.

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pAAVS1-BSD-DNR.dna
Map and Sequence File:    Download    Open   
Sequence Author:  OriGene
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KflI (7222) SexAI * (7122) AvrII (7111) XcmI (7042) PstI (6881) BsaI (6558) NsiI (6487) BbvCI (6452) AleI (6226) SacII (6140) FseI (6136) PmeI (6126) PfoI * (6094) PspXI (6048) NotI (6042) BsiWI (6036) MluI (6033) RsrII (6027) AscI - BssHII (5996) MreI - SgrAI (5992) AsiSI (5990) KpnI (5968) Acc65I (5964) AccI (5952) SalI (5951) EcoRI (5945) NcoI (5684) SnaBI (5664) NdeI (5558) BsmI (4837) BsrGI (4727) NruI * (4494) BsaBI * (4427) BspDI - ClaI (4323) ApaI (3756) PspOMI (3752) Bsu36I (7234) PciI (436) PsiI (2599) PmlI (3305) PshAI (3465) BstEII (3617) pAAVS1-BSD-DNR 7255 bp
KflI  (7222)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
SexAI  (7122)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
AvrII  (7111)
1 site
C C T A G G G G A T C C
XcmI  (7042)
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.
PstI  (6881)
1 site
C T G C A G G A C G T C
BsaI  (6558)
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.
NsiI  (6487)
1 site
A T G C A T T A C G T A
BbvCI  (6452)
1 site
C C T C A G C G G A G T C G
AleI  (6226)
1 site
C A C N N N N G T G G T G N N N N C A C
SacII  (6140)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
FseI  (6136)
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  (6126)
1 site
G T T T A A A C C A A A T T T G
PfoI  (6094)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
PspXI  (6048)
1 site
V C T C G A G B B G A G C T C V
NotI  (6042)
1 site
G C G G C C G C C G C C G G C G
BsiWI  (6036)
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.
MluI  (6033)
1 site
A C G C G T T G C G C A
RsrII  (6027)
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.
AscI  (5996)
1 site
G G C G C G C C C C G C G C G G
BssHII  (5996)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
MreI  (5992)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (5992)
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.
AsiSI  (5990)
1 site
G C G A T C G C C G C T A G C G
KpnI  (5968)
1 site
G G T A C C C C A T G G
Acc65I  (5964)
1 site
G G T A C C C C A T G G
AccI  (5952)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
SalI  (5951)
1 site
G T C G A C C A G C T G
EcoRI  (5945)
1 site
G A A T T C C T T A A G
NcoI  (5684)
1 site
C C A T G G G G T A C C
SnaBI  (5664)
1 site
T A C G T A A T G C A T
NdeI  (5558)
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.
BsmI  (4837)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
BsrGI  (4727)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
NruI  (4494)
1 site
T C G C G A A G C G C T
* Blocked by Dam methylation.
BsaBI  (4427)
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.
BspDI  (4323)
1 site
A T C G A T T A G C T A
ClaI  (4323)
1 site
A T C G A T T A G C T A
ApaI  (3756)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (3752)
1 site
G G G C C C C C C G G G
Bsu36I  (7234)
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.
PciI  (436)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PsiI  (2599)
1 site
T T A T A A A A T A T T
PmlI  (3305)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
PshAI  (3465)
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.
BstEII  (3617)
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.
AmpR
1256 .. 2116  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1256 .. 2047  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1256 .. 2116  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2048 .. 2116  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1256 .. 2116  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
497 .. 1085  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
497 .. 1085  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
HA-R
6720 .. 7249  =  530 bp
right homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene
HA-R
6720 .. 7249  =  530 bp
right homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene
HA-L
3278 .. 3779  =  502 bp
left homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene
HA-L
3278 .. 3779  =  502 bp
left homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene
PGK promoter
4768 .. 5267  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
4768 .. 5267  =  500 bp
mouse phosphoglycerate kinase 1 promoter
hGH poly(A) signal
6169 .. 6645  =  477 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
6169 .. 6645  =  477 bp
human growth hormone polyadenylation signal
f1 ori
2503 .. 2958  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
2503 .. 2958  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
4328 .. 4726  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
4328 .. 4726  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
CMV enhancer
5385 .. 5688  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
5385 .. 5688  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
5689 .. 5892  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
5689 .. 5892  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
AmpR promoter
2117 .. 2221  =  105 bp
AmpR promoter
2117 .. 2221  =  105 bp
MCS
5945 .. 6048  =  104 bp
multiple cloning site
MCS
5945 .. 6048  =  104 bp
multiple cloning site
Myc
6051 .. 6080  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
6051 .. 6080  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
FLAG
6099 .. 6122  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
6099 .. 6122  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
T7 promoter
5918 .. 5936  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
5918 .. 5936  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  2978 .. 3319  =  342 bp
ORF:  113 amino acids  =  13.1 kDa
ORF:  4424 .. 4822  =  399 bp
ORF:  132 amino acids  =  14.5 kDa
ORF:  1386 .. 1652  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  1256 .. 2116  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3257 .. 3571  =  315 bp
ORF:  104 amino acids  =  11.4 kDa
ORF:  4328 .. 4987  =  660 bp
ORF:  219 amino acids  =  23.0 kDa
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