LOCUS       sequence-392307-        6476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses V5-LOV-Turbo-NES in mammalian cells, AAV vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6476)
  AUTHORS   Lee SY, Cheah JS, Zhao B, Xu C, Roh H, Kim CK, Cho KF, Udeshi ND, 
            Carr SA, Ting AY
  TITLE     Engineered allostery in light-regulated LOV-Turbo enables precise 
            spatiotemporal control of proximity labeling in living cells.
  JOURNAL   Nat Methods. 2023 May 15. doi: 10.1038/s41592-023-01880-5.
  PUBMED    37188954
REFERENCE   2  (bases 1 to 6476)
  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. 2023 May 15. doi: 10.1038/s41592-023-01880-5."
COMMENT     Sequence Label: AAV_CAG_V5-LOV-Turbo-NES
FEATURES             Location/Qualifiers
     source          1..6476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        216..519
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        521..798
                     /label=chicken beta-actin promoter
     intron          799..1815
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1739..1761
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1826..1835
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1835..1876
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3257..3283
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(3364..3384)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3739..3879
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3739..3868
                     /label=AAV2 ITR
     rep_origin      3954..4409
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4041..4060)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4251..4272
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4426..4445)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4545..4567
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4605..4623)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4691..4795
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4796..4864,4865..5656)
                     /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(5014..5033)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5827..6415
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6316..6335
                     /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 gcgtctgcag gaatctagtt attaatagta atcaattacg
      181 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      241 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      301 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      361 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      421 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      481 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
      541 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
      601 taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc aggsggggsg
      661 gggsggggsg aggggsgggg sggggsgagg sggagaggtg cggcggcagc caatcagagc
      721 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
      781 cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc
      841 gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg
      901 cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct
      961 tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc
     1021 ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg
     1081 cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc
     1141 gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa
     1201 ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg
     1261 ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg
     1321 gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg
     1381 ggggtgccgg gcggggcggg gccgcctcgg gccggggrgg gctcgggggr ggggcgcggc
     1441 ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg
     1501 taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct
     1561 gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg
     1621 aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc
     1681 cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg
     1741 ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc
     1801 ttctttttcc tacagcaatt ggctagccac catgggcaag cccatcccca accccctgct
     1861 gggcctggac agcaccggta ccaaagacaa cacagtgcct ctgaagctga tcgccctcct
     1921 ggccaatgga gagttccaca gcggcgagca gctgggcgaa accctgggca tgagcagagc
     1981 cgctatcaac aagcacatcc agacccttcg cgactggggc gtcgacgtgt tcagcgtgcc
     2041 cggcaagggc tacagcctgc ctgagcctat ccccctgctg aacgccaagc agattcaggg
     2101 ccagctggaa tttagagcca ccaccctgga aagaatcgag aagtcttttg tgatcaccga
     2161 ccccagactg cctgataatc ctatcatctt cgtgtctgat agcttcctgc agctcaccga
     2221 gtattctcgg gaagagatcc tcggaagaaa ttgcagattc ctgcagggcc cagagacaga
     2281 tagagccaca gtgcggaaga tcagagatgc catcgacaac cagacagagg tgaccgtgca
     2341 gctgatcaac tacaccaagt ccggcaaaaa gttctggaac gttttccatc tccaacctat
     2401 gagagactac aagggcgacg tgcagtactt catcggagtt cagctggacg gcaccgagag
     2461 actgcacggc gccgcagagc gggaagctgt gtgcctggtc aagaagaccg ccttccagat
     2521 cgccgtggct gtgctgcccg tggtggacag cacaaaccag tacctgctgg acagaatcgg
     2581 cgagctgaag tccggcgacg cttgtatcgc cgaataccag caggccggca gaggcagcag
     2641 aggccggaaa tggttcagcc ccttcggcgc caacctgtac ctgagcatgt tttggcggct
     2701 gaaaagaggc cctgccgcca caggcctggg ccccgtgata ggaatcgtga tggccgaggc
     2761 tctgaggaag ctgggtgtgg acaaagtgcg ggtgaagtgg cctaacgacc tgtacctgca
     2821 ggacagaaag ctggccggca tcctggtgga actggctggc atcaccggcg acgccgctca
     2881 gattgtgatc ggcgctggca tcaatgtggc catgagacgc gtggaagaga gcgtggtcaa
     2941 ccagggctgg atcacactgc aagaggccgg tgtgaacctg gataggaaca ccctggccgc
     3001 caccctgatc agagagctga gagccgctct ggaattgttt gagcaggagg gcctggcccc
     3061 ttacctgcca cggtgggaga aactggataa cttcatcaac aggcctgtga agctgatcat
     3121 cggcgataag gaaatcctgg gaatcagccg gggcatcgac aagcagggag ccctgctgct
     3181 ggaacaagat ggagtgatta agccttggat gggcggagaa atcagcctgc ggtccgccga
     3241 gaagggatcc ctgcagctgc ctcccctgga gcgcctgacc ctggactaat agggccgaat
     3301 tcactagtat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3361 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3421 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac
     3481 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3541 tgacaattcc gtggtgttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3601 ctagctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     3661 ataaacaagt taacaacaac attgcattca ttttatgttt caggttcagg gggagatgtg
     3721 ggaactagtt gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3781 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     3841 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     3901 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     3961 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4021 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4081 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4141 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     4201 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4261 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     4321 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4381 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     4441 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4501 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4561 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     4621 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4681 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4741 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4801 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4861 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4921 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4981 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5041 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5101 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5161 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5221 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5281 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5341 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5401 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5461 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5521 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5581 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5641 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5701 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5761 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5821 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5881 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5941 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6001 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6061 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6121 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6181 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6241 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6301 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6361 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6421 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
