AAV pEF1a-DIO-FLPo-WPRE-hGHpA vector (V001291)

Price Information

Cat No. Plasmid Name Availability Add to cart
V001291 AAV pEF1a-DIO-FLPo-WPRE-hGHpA In stock (lyophilized plasmid)

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Two vials of lyophilized plasmid will be delivered, each vial is about 5µg.

Basic Vector Information

      • Vector Name:
      • AAV pEF1a-DIO-FLPo-WPRE-hGHpA
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 6892 bp
      • Type:
      • Mammalian Expression, AAV
      • Copy Number:
      • High Copy
      • Promoter:
      • EF1a
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • AGCTGACAGGTGGTGGCAAT
      • 3' Primer:
      • TCAAGCCTCAGACAGTGGTTC

AAV pEF1a-DIO-FLPo-WPRE-hGHpA vector Vector Map

AAV pEF1a-DIO-FLPo-WPRE-hGHpA6892 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600AAV2 ITR (alternate)EF-1-alpha promoterlox2272loxPFLPolox2272loxPWPREhGH poly(A) signalAAV2 ITRf1 oripRS-markerpGEX 3'pBRforEcoAmpR promoterAmpRori

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

AAV pEF1a-DIO-FLPo-WPRE-hGHpA vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6892 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  An AAV vector expresses FLPo in a Cre recombinase-dependent manner..
ACCESSION   .
VERSION     .
KEYWORDS    AAV pEF1a-DIO-FLPo-WPRE-hGHpA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6892)
  AUTHORS   Zingg B, Chou XL, Zhang ZG, Mesik L, Liang F, Tao HW, Zhang LI
  TITLE     AAV-Mediated Anterograde Transsynaptic Tagging: Mapping 
            Corticocollicular Input-Defined Neural Pathways for Defense 
            Behaviors.
  JOURNAL   Neuron. 2017 Jan 4;93(1):33-47. doi: 10.1016/j.neuron.2016.11.045. 
            Epub 2016 Dec 15.
  PUBMED    27989459
REFERENCE   2  (bases 1 to 6892)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2016.11.045"; journalName: "Neuron"; date: 
            "2017-01-4- 4"; volume: "93"; issue: "1"; pages: "33-47"
FEATURES             Location/Qualifiers
     source          1..6892
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   13..142
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        243..1421
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          474..1412
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1369..1389
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1452..1485
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    1536..1569
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1581..2879)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     protein_bind    complement(2889..2922)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    complement(2973..3006)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3031..3619
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3084..3104)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3502..3513)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3651..4127
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3850..3869)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4167..4307
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4167..4296
                     /label=AAV2 ITR
     rep_origin      4382..4837
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4469..4488)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4679..4700
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4854..4873)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4973..4995
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5033..5051)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5119..5223
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5224..6084
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5442..5461)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6255..6843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6744..6763
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"