LOCUS       sequence-292210-        7659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  can be used to generate AAV virus that will express light-inducible 
            site-specific iFlpV recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7659)
  AUTHORS   Yao S, Yuan P, Ouellette B, Zhou T, Mortrud M, Balaram P, Chatterjee
            S, Wang Y, Daigle TL, Tasic B, Kuang X, Gong H, Luo Q, Zeng S, 
            Curtright A, Dhaka A, Kahan A, Gradinaru V, Chrapkiewicz R, 
            Schnitzer M, Zeng H, Cetin A
  TITLE     RecV recombinase system for in vivo targeted optogenomic 
            modifications of single cells or cell populations.
  JOURNAL   Nat Methods. 2020 Mar 23. pii: 10.1038/s41592-020-0774-3. doi: 
            10.1038/s41592-020-0774-3.
  PUBMED    32203389
REFERENCE   2  (bases 1 to 7659)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2020 Mar 23. pii: 10.1038/s41592-020-0774-3. doi: 
            10.1038/s41592-020-0774-3."
COMMENT     Sequence Label: pAAV-EF1a-iFlpV
FEATURES             Location/Qualifiers
     source          1..7659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1986..2006
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2022..2078
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2088..2108
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    3786..4374
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3839..3859)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4257..4268)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4406..4882
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4605..4624)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4922..5062
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4922..5051
                     /label=AAV2 ITR
     rep_origin      5137..5592
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5224..5243)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5434..5455
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5609..5628)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5728..5750
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5788..5806)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5874..5978
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5979..6047,6048..6839)
                     /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(6197..6216)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7010..7598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7499..7518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc tgccgccacc atgagccagt
     1441 tcgacatcct gtgcaagacc ccccccaagg tgctggtgcg gcagttcgtg gagagattcg
     1501 agaggcccag cggtacccat acactgtatg cccctggagg atatgatatt atgggatatc
     1561 tgattcagat tatgaaaaga cctaatcctc aggtggaact gggacctgtg gatacaagcg
     1621 tggccctgat tctgtgtgat ctgaaacaga aagatacacc tattgtgtat gccagcgaag
     1681 cctttctgta tatgacagga tatagcaatg ccgaagtgct gggaagaaat tgtagatttc
     1741 tgcagagccc tgatggaatg gtgaaaccta aaagcacaag aaaatatgtg gatagcaata
     1801 caattaatac aatgagaaaa gccattgata gaaatgccga agtgcaggtg gaagtggtga
     1861 attttaaaaa aaatggacag agatttgtga attttctgac aatgattcct gtgagagatg
     1921 aaacaggaga atatagatat agcatgggat ttcagtgtga aacagaaggc ggaagcggtg
     1981 gcgtgcccaa gaagaagagg aaagtcggat ccggcagcgg cgccaccaac ttcagcctgc
     2041 tgaagcaggc cggcgacgtg gaggagaacc ccggccccct cgaggtgccc aagaagaaga
     2101 ggaaagtcgg cggacatact ctgtacgctc ccggagggta tgacataatg ggttacctaa
     2161 tccagattat gaagcgtcct aatccccagg ttgagttggg accagtggac accagcgtag
     2221 ccttaattct ttgcgacctt aaacagaaag atacaccaat agtatacgcg agtgaggctt
     2281 ttttgtatat gacgggttat tccaacgcgg aagtgttagg acgaaattgt cggtttctac
     2341 aatcaccgga tgggatggtc aaaccgaaaa gtactagaaa gtatgtggat tcgaatacaa
     2401 ttaacaccat gagaaaggca atcgatagga acgcagaagt acaagtcgag gttgtcaatt
     2461 tcaaaaagaa tgggcaacgc ttcgtaaact ttctcactat gatacctgtt agggacgaaa
     2521 cgggcgagta tcgatactct atgggcttcc aatgtgaaac agaaggtacc ggcgagaaga
     2581 tcgccagctg tgccgccgag ctgacctacc tgtgctggat gatcacccac aacggcaccg
     2641 ccatcaagag ggccaccttc atgagctaca acaccatcat cagcaacagc ctgagcttcg
     2701 acatcgtgaa caagagcctg cagttcaagt acaagaccca gaaggccacc atcctggagg
     2761 ccagcctgaa gaagctgatc cccgcctggg agttcaccat catcccttac aacggccaga
     2821 agcaccagag cgacatcacc gacatcgtgt ccagcctgca gctgcagttc gagagcagcg
     2881 aggaggccga caagggcaac agccacagca agaagatgct gaaggccctg ctgtccgagg
     2941 gcgagagcat ctgggagatc accgagaaga tcctgaacag cttcgagtac accagcaggt
     3001 tcaccaagac caagaccctg taccagttcc tgttcctggc cacattcatc aactgcggca
     3061 ggttcagcga catcaagaac gtggacccca agagcttcaa gctggtgcag aacaagtacc
     3121 tgggcgtgat cattcagtgc ctggtgaccg agaccaagac aagcgtgtcc aggcacatct
     3181 actttttcag cgccagaggc aggatcgacc ccctggtgta cctggacgag ttcctgagga
     3241 acagcgagcc cgtgctgaag agagtgaaca ggaccggcaa cagcagcagc aacaagcagg
     3301 agtaccagct gctgaaggac aacctggtgc gcagctacaa caaggccctg aagaagaacg
     3361 ccccctaccc catcttcgct atcaagaacg gccctaagag ccacatcggc aggcacctga
     3421 tgaccagctt tctgagcatg aagggcctga ccgagctgac aaacgtggtg ggcaactgga
     3481 gcgacaagag ggcctccgcc gtggccagga ccacctacac ccaccagatc accgccatcc
     3541 ccgaccacta cttcgccctg gtgtccaggt actacgccta cgaccccatc agcaaggaga
     3601 tgatcgccct gaaggacgag accaacccca tcgaggagtg gcagcacatc gagcagctga
     3661 agggcagcgc cgagggcagc atcagatacc ccgcctggaa cggcatcatc agccaggagg
     3721 tgctggacta cctgagcagc tacatcaaca ggcggatctg agaattcgat atcaagctta
     3781 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     3841 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     3901 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     3961 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     4021 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     4081 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     4141 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     4201 tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     4261 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     4321 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     4381 accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct
     4441 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     4501 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     4561 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     4621 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     4681 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     4741 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     4801 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     4861 cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg
     4921 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     4981 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     5041 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     5101 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     5161 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     5221 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5281 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5341 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5401 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5461 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     5521 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5581 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5641 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5701 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5761 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5821 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5881 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5941 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     6001 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6061 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     6121 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6181 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6241 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6301 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6361 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6421 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6481 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     6541 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6601 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6661 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6721 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6781 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6841 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6901 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6961 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7021 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7081 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     7141 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7201 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7261 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7321 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7381 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7441 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7501 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7561 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7621 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
