pAAV2-EF1a-tGFP-WPRE

Adeno-associated virus (AAV) vector for expressing a C-terminally TurboGFP-tagged protein using the EF-1a promoter.

Sequence Author: OriGene

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HomePlasmidsViral Expression & Packaging VectorspAAV2-EF1a-tGFP-WPRE
AlwNI (5812) FspI (5110) ScaI (4852) PsiI (4062) RsrII (3474) BstEII (3461) BstXI (3407) NsiI (3291) SphI (3289) NdeI (390) MfeI (471) AgeI (478) AflII (779) PasI (1147) BstAPI (1179) Eco53kI (1225) SacI (1227) PaeR7I - XhoI (1364) EF51-F sequencing primer (1526 .. 1545) Acc65I (1578) KpnI (1582) BspDI * - ClaI * (1591) AscI (1621) MluI (1628) SalI (1634) AccI (1635) HincII (1636) BspQI - SapI (1769) AC_GFP-R sequencing primer (1748 .. 1769) PaqCI (2091) BsaBI * (2103) PmeI (2342) AleI (3030) pAAV2-EF1a-tGFP-WPRE 6226 bp
AlwNI  (5812)
1 site		 C A G N N N C T G G T C N N N G A C
Sticky ends from different AlwNI sites may not be compatible.
FspI  (5110)
1 site		 T G C G C A A C G C G T
ScaI  (4852)
1 site		 A G T A C T T C A T G A
PsiI  (4062)
1 site		 T T A T A A A A T A T T
RsrII  (3474)
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.
BstEII  (3461)
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.
BstXI  (3407)
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.
NsiI  (3291)
1 site		 A T G C A T T A C G T A
SphI  (3289)
1 site		 G C A T G C C G T A C G
NdeI  (390)
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.
MfeI  (471)
1 site		 C A A T T G G T T A A C
AgeI  (478)
1 site		 A C C G G T T G G C C A
AflII  (779)
1 site		 C T T A A G G A A T T C
PasI  (1147)
1 site		 C C C W G G G G G G W C C C
Sticky ends from different PasI sites may not be compatible.
BstAPI  (1179)
1 site		 G C A N N N N N T G C C G T N N N N N A C G
Sticky ends from different BstAPI sites may not be compatible.
Eco53kI  (1225)
1 site		 G A G C T C C T C G A G
SacI  (1227)
1 site		 G A G C T C C T C G A G
PaeR7I  (1364)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1364)
1 site		 C T C G A G G A G C T C
Acc65I  (1578)
1 site		 G G T A C C C C A T G G
KpnI  (1582)
1 site		 G G T A C C C C A T G G
BspDI  (1591)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1591)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
AscI  (1621)
1 site		 G G C G C G C C C C G C G C G G
MluI  (1628)
1 site		 A C G C G T T G C G C A
SalI  (1634)
1 site		 G T C G A C C A G C T G
AccI  (1635)
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.
HincII  (1636)
1 site		 G T Y R A C C A R Y T G
BspQI  (1769)
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  (1769)
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.
PaqCI  (2091)
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.
BsaBI  (2103)
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.
PmeI  (2342)
1 site		 G T T T A A A C C A A A T T T G
AleI  (3030)
1 site		 C A C N N N N G T G G T G N N N N C A C
EF51-F sequencing primer
20-mer  /  55% GC
1 binding site
1526 .. 1545  =  20 annealed bases
Tm  =  58°C
AC_GFP-R sequencing primer
22-mer  /  64% GC
1 binding site
1748 .. 1769  =  22 annealed bases
Tm  =  66°C
EF-1α promoter
400 .. 1577  =  1178 bp
strong constitutive promoter for human elongation factor EF-1α
EF-1α promoter
400 .. 1577  =  1178 bp
strong constitutive promoter for human elongation factor EF-1α
AmpR
4546 .. 5406  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4546 .. 4614  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4546 .. 5406  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4615 .. 5406  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4546 .. 5406  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
TurboGFP
1640 .. 2335  =  696 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
TurboGFP
1640 .. 2335  =  696 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
WPRE
2361 .. 2949  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
2361 .. 2949  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
5577 .. 6165  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5577 .. 6165  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
hGH poly(A) signal
2973 .. 3449  =  477 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
2973 .. 3449  =  477 bp
human growth hormone polyadenylation signal
f1 ori
3704 .. 4159  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3704 .. 4159  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
AAV2 ITR
1 .. 141  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AAV2 ITR
1 .. 141  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AAV2 ITR
3489 .. 3629  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AAV2 ITR
3489 .. 3629  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AmpR promoter
4441 .. 4545  =  105 bp
AmpR promoter
4441 .. 4545  =  105 bp
MCS
1578 .. 1639  =  62 bp
multiple cloning site
MCS
1578 .. 1639  =  62 bp
multiple cloning site
EF-1α intron A
631 .. 1568  =  938 bp
intron upstream of the start codon of human EF-1α
EF-1α intron A
631 .. 1568  =  938 bp
intron upstream of the start codon of human EF-1α
ORF:  2464 .. 3054  =  591 bp
ORF:  196 amino acids  =  20.8 kDa
ORF:  4546 .. 5406  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  956 .. 1267  =  312 bp
ORF:  103 amino acids  =  10.7 kDa
ORF:  1640 .. 2344  =  705 bp
ORF:  234 amino acids  =  25.9 kDa
ORF:  3287 .. 3727  =  441 bp
ORF:  146 amino acids  =  16.0 kDa
ORF:  2112 .. 3086  =  975 bp
ORF:  324 amino acids  =  35.0 kDa
ORF:  698 .. 1036  =  339 bp
ORF:  112 amino acids  =  13.0 kDa
ORF:  892 .. 1317  =  426 bp
ORF:  141 amino acids  =  15.5 kDa
ORF:  1525 .. 1761  =  237 bp
ORF:  78 amino acids  =  8.7 kDa
ORF:  3337 .. 3684  =  348 bp
ORF:  115 amino acids  =  13.4 kDa
ORF:  2757 .. 2981  =  225 bp
ORF:  74 amino acids  =  8.1 kDa
ORF:  5010 .. 5276  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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Individual Sequences & Maps

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pLVX-rHom-Nuc1
pRetroX-TetOne
pAAV-AC-RFP
pLenti6.3 TO V5-DEST
pLVX-rHom-Sec1
pRetroX-TetOne-Puro
pAAV2-EF1a-tGFP-WPRE
pLenti6.3 V5-DEST
pLVX-Tet-Off Advanced
pRetroX-Tight-Hyg
pAd BLOCK-iT-DEST
pLenti6.4 R4R2 V5-DEST
pLVX-Tet-On Advanced
pRetroX-Tight-Pur
pAd CMV V5-DEST
pLenti6 BLOCK-iT-DEST
pLVX-Tet3G
pRetroX-TRE3G
pAd PL-DEST
pLenti6 capTEV-CT-DEST
pLVX-TetOne
pRFP-C-RS
pAdenoX-CMV Linear
pLenti6 capTEV-NT-DEST1
pLVX-TetOne-Puro
pRFP-CB-shLenti
pAdenoX-DsRed-Express Linear
pLenti6 TR
pLVX-Tight-Puro
pRS
pAdenoX-PRLS Linear
pLenti6 UbC V5-DEST
pLVX-TRE3G
pRSGC1-U6-sg-HTS6-CMV-Cas9-2A-Puro
pAdenoX-PRLS-DsRed-Express Linear
pLenti6 V5-DEST
pLVX-TRE3G-Hom1
pRSGC2-U6-sg-HTS6-UbiC-Cas9-2A-Puro
pAdenoX-PRLS-ZsGreen1 Linear
pLenti7.3 V5-DEST
pLVX-TRE3G-IRES
pRSI12-U6-sh-HTS4-UbiC-TagRFP-2A-Puro
pAdenoX-Tet3G Linear
pLEX-MCS
pLVX-TRE3G-mCherry
pRSI16-U6-sh-HTS6-UbiC-TagRFP-2A-Puro
pAdenoX-ZsGreen1 Linear
pLHCX
pLVX-TRE3G-ZsGreen1
pRSI16cb-U6-sh-13kCB18-HTS6-UbiC-TagRFP-2A-Puro
pB-RS
pLKO.1
pLX301
pRSI17-U6-sh-HTS6-UbiC-TagGFP2-2A-Puro
pBABE-Hygro
pLKO.1 puro
pLX302
pRSI17cb-U6-sh-13kCB18-HTS6-UbiC-TagGFP2-2A-Puro
pBABE-Neo
pLMN-ZsGreen-Neomycin
pLX303
pRSIT12-U6Tet-sh-HTS4-CMV-TetRep-2A-TagRFP-2A-Puro
pBABE-Puro
pLMPd-Ametrine
pLX304
pRSIT16-U6Tet-sh-HTS6-CMV-TetRep-2A-TagRFP-2A-Puro
pBABE-Zeo
pLNCX2
pLX304-V5-Blast-Empty
pRSIT17-U6Tet-sh-HTS6-CMV-TetRep-2A-TagGFP2-2A-Puro
pCMV-VSV-G
pLP VSVG
pLXIN
pRSITEP-U6Tet-sh-noHTS-EF1-TetRep-2A-Puro
pGFP-B-RS
pLP1
pLXSN
pRSITUR-U6Tet-sh-HTS3-UbiC-TetRep-2A-TagRFP
pGFP-C-shLenti
pLP2
pMD2.G
pShuttle2
pGFP-V-RS
pLPCX
pMSCV16URP-U6-sh-HTS6-UbiC-TagRFP-2A-Puro
pSIREN-RetroQ
pGIPZ
pLVX-EF1alpha-IRES-mCherry
pMSCVhyg
pSIREN-RetroQ-DsRed-Express
pLenti-C-GFP
pLVX-EF1alpha-IRES-Puro
pMSCVneo
pSIREN-RetroQ-TetH
pLenti-C-mGFP
pLVX-EF1alpha-IRES-ZsGreen1
pMSCVpuro
pSIREN-RetroQ-TetP
pLenti-C-Myc-DDK
pLVX-EF1alpha-Tet3G
pQC-tTS-IN
pSIREN-RetroQ-ZsGreen1
pLenti-C-Myc-DDK-IRES-BSD
pLVX-Het-1
pQCXIH
psPAX2
pLenti-C-Myc-DDK-IRES-GFP
pLVX-Het-2
pQCXIN
pTRE3G-Hom1
pLenti-C-Myc-DDK-IRES-Neo
pLVX-Het-Mem1
pQCXIP
pTRIPZ
pLenti-C-Myc-DDK-IRES-Puro
pLVX-Het-Nuc1
pQCXIX
ptTS-Neo
pLenti-C-Myc-DDK-IRES-tRFP
pLVX-Hom-1
pRetro-Lib
pZIP-hCMV-RFP-Puro
pLenti-C-tRFP
pLVX-Hom-Mem1
pRetroQ-AcGFP1-C1
pZIP-hCMV-ZSGreen-Puro
pLenti-C-tYFP
pLVX-Hom-Nuc1
pRetroQ-AcGFP1-N1
pZIP-hEF1-alpha-RFP-Puro
pLenti-EF1a-C-mGFP
pLVX-IRES-Hyg
pRetroQ-DsRed-Monomer-C1
pZIP-hEF1-alpha-ZsGreen-Puro
pLenti-EF1a-C-Myc-DDK-IRES-Puro
pLVX-IRES-mCherry
pRetroQ-DsRed-Monomer-N1
pZIP-mCMV-RFP-Puro
pLenti-EF1a-C-tGFP
pLVX-IRES-Neo
pRetroQ-mCherry-C1
pZIP-mCMV-ZsGreen-Puro
pLenti-N-tGFP
pLVX-IRES-Puro
pRetroQ-mCherry-N1
pZIP-mEF1-alpha-RFP-Puro
pLenti-N-tRFP
pLVX-IRES-tdTomato
pRetroX-IRES-DsRedExpress
pZIP-mEF1-alpha-ZsGreen-Puro
pLenti-N-tYFP
pLVX-IRES-ZsGreen1
pRetroX-IRES-ZsGreen1
pZIP-SFFV-RFP-Puro
pLenti3.3 TR
pLVX-PTuner
pRetroX-PTuner
pZIP-SFFV-ZsGreen-Puro
pLenti4 BLOCK-iT-DEST
pLVX-PTuner Green
pRetroX-PTuner IRES
pZIP-TRE3G-ZsGreen-Puro
pLenti4 V5-DEST
pLVX-PTunerC
pRetroX-Tet-Off Advanced
pSIREN-Shuttle (linearized)
pLenti6.2 C-Lumio V5-DEST
pLVX-Puro
pRetroX-Tet-On Advanced
scramble shRNA

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