LOCUS       sequence-292209-        7443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  can be used to generate AAV virus that will express light-inducible 
            site-specific iDreV recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7443)
  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 7443)
  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-iDreV
FEATURES             Location/Qualifiers
     source          1..7443
                     /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             1440..1460
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2112..2132
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2148..2204
                     /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             2214..2234
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    3570..4158
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3623..3643)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4041..4052)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4190..4666
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4389..4408)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4706..4846
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4706..4835
                     /label=AAV2 ITR
     rep_origin      4921..5376
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5008..5027)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5218..5239
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5393..5412)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5512..5534
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5572..5590)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5658..5762
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5763..5831,5832..6623)
                     /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(5981..6000)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6794..7382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7283..7302
                     /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 accatggccc
     1441 ctaagaagaa gaggaaggtt tctgagctga ttattagtgg ttcatctggt ggattcctgc
     1501 gaaacatcgg caaagagtat caggaggccg ctgaaaactt catgaggttt atgaatgacc
     1561 agggggcgta cgctcctaac actttgaggg atttgaggtt ggtctttcat agctgggcca
     1621 gatggtgcca tgctcggggt acccatacac tgtatgcccc tggaggatat gatattatgg
     1681 gatatctgat tcagattatg aaaagaccta atcctcaggt ggaactggga cctgtggata
     1741 caagcgtggc cctgattctg tgtgatctga aacagaaaga tacacctatt gtgtatgcca
     1801 gcgaagcctt tctgtatatg acaggatata gcaatgccga agtgctggga agaaattgta
     1861 gatttctgca gagccctgat ggaatggtga aacctaaaag cacaagaaaa tatgtggata
     1921 gcaatacaat taatacaatg agaaaagcca ttgatagaaa tgccgaagtg caggtggaag
     1981 tggtgaattt taaaaaaaat ggacagagat ttgtgaattt tctgacaatg attcctgtga
     2041 gagatgaaac aggagaatat agatatagca tgggatttca gtgtgaaaca gaaggcggaa
     2101 gcggtggcgt gcccaagaag aagaggaaag tcggatccgg cagcggcgcc accaacttca
     2161 gcctgctgaa gcaggccggc gacgtggagg agaaccccgg ccccctcgag gtgcccaaga
     2221 agaagaggaa agtcggcgga catactctgt acgctcccgg agggtatgac ataatgggtt
     2281 acctaatcca gattatgaag cgtcctaatc cccaggttga gttgggacca gtggacacca
     2341 gcgtagcctt aattctttgc gaccttaaac agaaagatac accaatagta tacgcgagtg
     2401 aggctttttt gtatatgacg ggttattcca acgcggaagt gttaggacga aattgtcggt
     2461 ttctacaatc accggatggg atggtcaaac cgaaaagtac tagaaagtat gtggattcga
     2521 atacaattaa caccatgaga aaggcaatcg ataggaacgc agaagtacaa gtcgaggttg
     2581 tcaatttcaa aaagaatggg caacgcttcg taaactttct cactatgata cctgttaggg
     2641 acgaaacggg cgagtatcga tactctatgg gcttccaatg tgaaacagaa ggtacccagc
     2701 ttgcatggtt tccaattagt cctgaaatgg cacgcgaata ctttcttcag ttgcacgatg
     2761 cagacctggc ctccactacc atcgacaagc actatgctat gcttaatatg cttctgtccc
     2821 actgcggact gccacccttg tccgacgaca agtcagtgag tcttgccatg agaagaatta
     2881 gaagagaagc cgcaaccgaa aagggtgaga ggacaggaca ggcaatcccc ctgcgctggg
     2941 acgacctgaa gctgctggat gtgctgctca gcaggagcga gcggctggtc gacctgcgca
     3001 acagggcttt cctgttcgta gcctataaca ccctcatgag aatgtctgaa atatcacgca
     3061 tcagggttgg ggacttggat cagacaggag acacagtgac cctgcacatc agtcacacta
     3121 agacaatcac cacagctgcg ggccttgaca aagtgctctc ccggcgaacc acagcagtgc
     3181 tcaatgactg gctggacgtc agtgggctta gagaacatcc agacgctgtg ctcttcccac
     3241 ctatacaccg gtcaaacaaa gcccgcatta ctaccacgcc cctgaccgcc cctgccatgg
     3301 agaagatttt cagtgatgcc tgggtgctgc tgaacaaacg ggacgccacc cccaataaag
     3361 ggaggtatag gacctggacc ggccattccg ccagggtggg tgccgcaata gacatggccg
     3421 agaaacaggt gtctatggtc gagattatgc aggaagggac atggaagaag cctgaaacac
     3481 tgatgcggta tctcagaagg ggcggagtgt ccgtgggagc caattctcga ctgatggata
     3541 gctaagaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     3601 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3661 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3721 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3781 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3841 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3901 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3961 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga
     4021 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     4081 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     4141 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc
     4201 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     4261 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     4321 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     4381 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     4441 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     4501 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     4561 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     4621 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     4681 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     4741 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     4801 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     4861 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     4921 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4981 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     5041 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     5101 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     5161 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     5221 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     5281 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     5341 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     5401 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     5461 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     5521 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     5581 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     5641 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5701 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5761 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5821 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5881 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5941 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6001 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6061 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6121 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6181 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6241 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6301 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6361 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6421 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6481 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6541 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6601 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6661 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6721 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6781 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6841 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6901 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6961 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7021 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7081 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7141 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7201 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7261 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7321 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7381 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7441 tgt
//
