pAAV-AC-GFP

Adeno-associated virus (AAV) vector for expressing a C-terminally TurboGFP-tagged protein using the CMV promoter.

Sequence Author: OriGene

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PciI (5712) FspI (4601) ScaI (4343) PfoI (3785) PsiI (3553) DraIII (3428) SbfI (3118) BstEII (2952) NsiI (2782) BsrGI (432) HincII (473) NdeI (723) Eco53kI (1055) SacI (1057) SacII (1144) BGI-F sequencing primer (1506 .. 1525) BspDI * - ClaI * (1650) AsiSI (1677) MreI - SgrAI (1679) AscI (1683) HindIII (1697) MluI (1720) BsiWI (1723) PaeR7I - XhoI (1735) AC_GFP-R sequencing primer (1849 .. 1870) PflMI (1926) PaqCI (2192) PspOMI (2325) ApaI (2329) PmeI (2443) AleI (2521) BbvCI - Bpu10I (2747) SphI (2780) pAAV-AC-GFP 5725 bp
PciI  (5712)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
FspI  (4601)
1 site
T G C G C A A C G C G T
ScaI  (4343)
1 site
A G T A C T T C A T G A
PfoI  (3785)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
PsiI  (3553)
1 site
T T A T A A A A T A T T
DraIII  (3428)
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.
SbfI  (3118)
1 site
C C T G C A G G G G A C G T C C
BstEII  (2952)
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.
NsiI  (2782)
1 site
A T G C A T T A C G T A
BsrGI  (432)
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.
HincII  (473)
1 site
G T Y R A C C A R Y T G
NdeI  (723)
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.
Eco53kI  (1055)
1 site
G A G C T C C T C G A G
SacI  (1057)
1 site
G A G C T C C T C G A G
SacII  (1144)
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.
BspDI  (1650)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1650)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
AsiSI  (1677)
1 site
G C G A T C G C C G C T A G C G
MreI  (1679)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (1679)
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.
AscI  (1683)
1 site
G G C G C G C C C C G C G C G G
HindIII  (1697)
1 site
A A G C T T T T C G A A
MluI  (1720)
1 site
A C G C G T T G C G C A
BsiWI  (1723)
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.
PaeR7I  (1735)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1735)
1 site
C T C G A G G A G C T C
PflMI  (1926)
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.
PaqCI  (2192)
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 PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
PspOMI  (2325)
1 site
G G G C C C C C C G G G
ApaI  (2329)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PmeI  (2443)
1 site
G T T T A A A C C A A A T T T G
AleI  (2521)
1 site
C A C N N N N G T G G T G N N N N C A C
BbvCI  (2747)
1 site
C C T C A G C G G A G T C G
Bpu10I  (2747)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
SphI  (2780)
1 site
G C A T G C C G T A C G
BGI-F sequencing primer
20-mer  /  45% GC
1 binding site
1506 .. 1525  =  20 annealed bases
Tm  =  53°C
AC_GFP-R sequencing primer
22-mer  /  64% GC
1 binding site
1849 .. 1870  =  22 annealed bases
Tm  =  66°C
AmpR
4037 .. 4897  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4037 .. 4105  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4037 .. 4897  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4106 .. 4897  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4037 .. 4897  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
TurboGFP
1741 .. 2436  =  696 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
TurboGFP
1741 .. 2436  =  696 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
ori
5068 .. 5656  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5068 .. 5656  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
hGH poly(A) signal
2464 .. 2940  =  477 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
2464 .. 2940  =  477 bp
human growth hormone polyadenylation signal
f1 ori
3195 .. 3650  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3195 .. 3650  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
474 .. 853  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
474 .. 853  =  380 bp
human cytomegalovirus immediate early enhancer
chimeric intron
1206 .. 1584  =  379 bp
fusion between human cytomegalovirus intron A and human β-globin intron 2
chimeric intron
1206 .. 1584  =  379 bp
fusion between human cytomegalovirus intron A and human β-globin intron 2
CMV promoter
854 .. 1057  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
854 .. 1057  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
AAV2 ITR
2980 .. 3120  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AAV2 ITR
2980 .. 3120  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AmpR promoter
3932 .. 4036  =  105 bp
AmpR promoter
3932 .. 4036  =  105 bp
MCS
1673 .. 1740  =  68 bp
multiple cloning site
MCS
1673 .. 1740  =  68 bp
multiple cloning site
ORF:  1741 .. 2445  =  705 bp
ORF:  234 amino acids  =  25.9 kDa
ORF:  2213 .. 2545  =  333 bp
ORF:  110 amino acids  =  11.7 kDa
ORF:  4037 .. 4897  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2778 .. 3218  =  441 bp
ORF:  146 amino acids  =  16.0 kDa
ORF:  32 .. 283  =  252 bp
ORF:  83 amino acids  =  9.0 kDa
ORF:  2828 .. 3175  =  348 bp
ORF:  115 amino acids  =  13.4 kDa
ORF:  1705 .. 2472  =  768 bp
ORF:  255 amino acids  =  25.1 kDa
ORF:  4501 .. 4767  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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