LOCUS       sequence-292208-        7374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Can be used to generate AAV virus that will express light-inducible 
            site-specific iCreV recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7374)
  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 7374)
  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-iCreV
FEATURES             Location/Qualifiers
     source          1..7374
                     /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"
     regulatory      1428..1437
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2037..2057
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2073..2129
                     /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             2139..2159
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    3501..4089
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3554..3574)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3972..3983)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4121..4597
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4320..4339)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4637..4777
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4637..4766
                     /label=AAV2 ITR
     rep_origin      4852..5307
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4939..4958)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5149..5170
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5324..5343)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5443..5465
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5503..5521)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5589..5693
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5694..5762,5763..6554)
                     /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(5912..5931)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6725..7313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7214..7233
                     /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 ttaagccgcc accatggcca
     1441 cctctgatga agtcaggaag aacctgatgg acatgttcag ggacaggcag gccttctctg
     1501 aacacacctg gaagatgctc ctgtctgtgt gcagatcctg ggctgcctgg tgcaagctga
     1561 acggtaccca tacactgtat gcccctggag gatatgatat tatgggatat ctgattcaga
     1621 ttatgaaaag acctaatcct caggtggaac tgggacctgt ggatacaagc gtggccctga
     1681 ttctgtgtga tctgaaacag aaagatacac ctattgtgta tgccagcgaa gcctttctgt
     1741 atatgacagg atatagcaat gccgaagtgc tgggaagaaa ttgtagattt ctgcagagcc
     1801 ctgatggaat ggtgaaacct aaaagcacaa gaaaatatgt ggatagcaat acaattaata
     1861 caatgagaaa agccattgat agaaatgccg aagtgcaggt ggaagtggtg aattttaaaa
     1921 aaaatggaca gagatttgtg aattttctga caatgattcc tgtgagagat gaaacaggag
     1981 aatatagata tagcatggga tttcagtgtg aaacagaagg cggaagcggt ggcgtgccca
     2041 agaagaagag gaaagtcgga tccggcagcg gcgccaccaa cttcagcctg ctgaagcagg
     2101 ccggcgacgt ggaggagaac cccggccccc tcgaggtgcc caagaagaag aggaaagtcg
     2161 gcggacatac tctgtacgct cccggagggt atgacataat gggttaccta atccagatta
     2221 tgaagcgtcc taatccccag gttgagttgg gaccagtgga caccagcgta gccttaattc
     2281 tttgcgacct taaacagaaa gatacaccaa tagtatacgc gagtgaggct tttttgtata
     2341 tgacgggtta ttccaacgcg gaagtgttag gacgaaattg tcggtttcta caatcaccgg
     2401 atgggatggt caaaccgaaa agtactagaa agtatgtgga ttcgaataca attaacacca
     2461 tgagaaaggc aatcgatagg aacgcagaag tacaagtcga ggttgtcaat ttcaaaaaga
     2521 atgggcaacg cttcgtaaac tttctcacta tgatacctgt tagggacgaa acgggcgagt
     2581 atcgatactc tatgggcttc caatgtgaaa cagaaggtac caacaggaaa tggttccctg
     2641 ctgaacctga ggatgtgagg gactacctcc tgtacctgca agccagaggc ctggctgtga
     2701 agaccatcca acagcacctg ggccagctca acatgctgca caggagatcc ggcctgcctc
     2761 gcccttctga ctccaatgct gtgtccctgg tgatgaggag aatcagaaag gagaatgtgg
     2821 atgctgggga gagagccaag caggccctgg cctttgaacg cactgacttt gaccaagtca
     2881 gatccctgat ggagaactct gacagatgcc aggacatcag gaacctggcc ttcctgggca
     2941 ttgcctacaa caccctgctg cgcattgccg aaattgccag aatcagagtg aaggacatct
     3001 cccgcaccga tggtgggaga atgctgatcc acattggcag gaccaagacc ctggtgtcca
     3061 cagctggtgt ggagaaggcc ctgtccctgg gggttaccaa gctggtggag agatggatct
     3121 ctgtgtctgg tgtggctgat gaccccaaca actacctgtt ctgccgggtc agaaagaatg
     3181 gtgtggctgc cccttctgcc acctcccaac tgtccacccg ggccctggaa gggatctttg
     3241 aggccaccca ccgcctgatc tatggtgcca aggatgactc tgggcagaga tacctggcct
     3301 ggtctggcca ctctgccaga gtgggtgctg ccagggacat ggccagggct ggtgtgtcca
     3361 tccctgaaat catgcaggct ggtggctgga ccaatgtgaa cattgtgatg aactacatca
     3421 gaaacctgga ctctgagact ggggccatgg tgaggctgct cgaagatggg gactaagaat
     3481 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     3541 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     3601 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     3661 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     3721 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     3781 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     3841 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     3901 cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct
     3961 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     4021 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     4081 cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc
     4141 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
     4201 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
     4261 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     4321 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     4381 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
     4441 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
     4501 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
     4561 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg
     4621 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     4681 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     4741 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     4801 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     4861 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4921 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4981 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     5041 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     5101 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5161 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     5221 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5281 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     5341 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     5401 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     5461 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     5521 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     5581 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5641 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5701 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5761 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5821 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5881 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5941 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     6001 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     6061 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     6121 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     6181 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     6241 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     6301 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6361 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6421 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6481 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6541 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6601 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6661 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6721 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6781 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6841 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6901 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6961 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     7021 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     7081 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     7141 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     7201 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     7261 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7321 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
