LOCUS       sequence-292202-        5934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  can be used to generate AAV virus that will express fusion protein 
            of split Cre (N-terminal) and fungal photoreceptor Vivid (VVD) 
            monomer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5934)
  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 5934)
  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-NCreV
FEATURES             Location/Qualifiers
     source          1..5934
                     /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"
     misc_feature    2061..2649
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2114..2134)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2532..2543)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2681..3157
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2880..2899)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3197..3337
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3197..3326
                     /label=AAV2 ITR
     rep_origin      3412..3867
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3499..3518)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3709..3730
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3884..3903)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4003..4025
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4063..4081)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4149..4253
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4254..4322,4323..5114)
                     /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(4472..4491)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5285..5873
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5774..5793
                     /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 ctctagagtc gacgccgcca
     1441 ccatgacgag tgatgaggtt cgcaagaacc tgatggacat gttcagggat cgccaggcgt
     1501 tttctgagca tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg gcatggtgca
     1561 agttgaatgg aggcggtgga ggtcatacac tgtatgcccc tggaggatat gatattatgg
     1621 gatatctgat tcagattatg aaaagaccta atcctcaggt ggaactggga cctgtggata
     1681 caagcgtggc cctgattctg tgtgatctga aacagaaaga tacacctatt gtgtatgcca
     1741 gcgaagcctt tctgtatatg acaggatata gcaatgccga agtgctggga agaaattgta
     1801 gatttctgca gagccctgat ggaatggtga aacctaaaag cacaagaaaa tatgtggata
     1861 gcaatacaat taatacaatg agaaaagcca ttgatagaaa tgccgaagtg caggtggaag
     1921 tggtgaattt taaaaaaaat ggacagagat ttgtgaattt tctgacaatg attcctgtga
     1981 gagatgaaac aggagaatat agatatagca tgggatttca gtgtgaaaca gaataagaat
     2041 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     2101 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2161 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2221 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     2281 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     2341 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     2401 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     2461 cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct
     2521 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     2581 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     2641 cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc
     2701 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
     2761 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
     2821 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     2881 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     2941 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
     3001 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
     3061 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
     3121 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg
     3181 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3241 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3301 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     3361 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     3421 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3481 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3541 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3601 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3661 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3721 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     3781 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3841 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     3901 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3961 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4021 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4081 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4141 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4201 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4261 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4321 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4381 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4441 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4501 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4561 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4621 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4681 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4741 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4801 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4861 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4921 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4981 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5041 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5101 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5161 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5221 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5281 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5341 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5401 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5461 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5521 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5581 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5641 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5701 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5761 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5821 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5881 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
