LOCUS       sequence-315078-        5802 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  optoNES construct with modified mTurquoise2-LOV2 RET.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5802)
  AUTHORS   Li LL, Klein FM, Li Greci L, Popinigis A, Freudenberg F, Courtney MJ
  TITLE     Resonance energy transfer sensitises and monitors in situ switching 
            of LOV2-based optogenetic actuators.
  JOURNAL   Nat Commun. 2020 Oct 9;11(1):5107. doi: 10.1038/s41467-020-18816-8.
  PUBMED    33037199
REFERENCE   2  (bases 1 to 5802)
  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 
            Commun."; date: "2020-10-9"; volume: "11(1):5107. doi"; pages: " 
            10.1038/s41467-020-18816-8"
COMMENT     Sequence Label: 
            AAV-CMV-NLS-mTurquoise2dC11-408LOV2-Ja(delta)-NES21-WPRE
FEATURES             Location/Qualifiers
     source          1..5802
                     /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        205..508
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        509..712
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     663..683
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      742..751
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(787..789,790..1302)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIKA
                     NFKIRHNIE"
     primer_bind     complement(829..850)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1090..1109)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1437..1458
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1931..2519
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1984..2004)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2402..2413)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2549..3025
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2748..2767)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3065..3205
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3065..3194
                     /label=AAV2 ITR
     rep_origin      3280..3735
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3367..3386)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3577..3598
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3752..3771)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3871..3893
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3931..3949)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4017..4121
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4122..4190,4191..4982)
                     /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(4340..4359)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5153..5741
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5642..5661
                     /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 gcgtgtgtct agaatagtaa tcaattacgg ggtcattagt
      181 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      241 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      301 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      361 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      421 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      481 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      541 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      601 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      661 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      721 gaaccgtcag atccgctagc cgccaccatg gccgcagcaa agcgcgtgaa gctggaccta
      781 ccggtcgtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      841 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      901 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      961 accctcgtga ccaccctgtc ctggggcgtg cagtgcttcg cccgctaccc cgaccacatg
     1021 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1081 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1141 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1201 cacaagctgg agtacaacta ctttagcgac aacgtctata tcaccgccga caagcagaag
     1261 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
     1321 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1381 cactacctga gcacccagtc caagcttagc aaagacccca acgagaagcg cgatcacatg
     1441 gtcctgctgg agttcgtgac cgccgccctc gagagaatcg agaaaaactt cgtcatcacc
     1501 gaccctagac tgcccgataa tcctatcatc ttcgcctccg actcatttct ccagctcact
     1561 gagtactctc gggaggaaat tctgggccgg aattgccggt tcctccaggg acctgaaacc
     1621 gatagagcca ctgtccgaaa aatccgggat gccatcgaca atcagactga ggtgaccgtc
     1681 cagctcatca attacaccaa atctggcaaa aaattctgga acctgttcca cctccagcct
     1741 atgagagatc agaagggcga cgtccagtac tttattggcg tccagctgga tggaactgaa
     1801 catgtgaggg acgctgcaga acgagagggc gtgatgctca tcaaaaaaac cgccgaaaac
     1861 atcgatgagc tgctgaagga gctggccgac ctgaacctgg actagggatc cgcggccgct
     1921 agataactga aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     1981 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2041 tgcttcccgt atggccttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2101 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2161 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2221 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2281 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     2341 ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     2401 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2461 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     2521 tcgattgtac cgcgggcccg ggatctacgg gtggcatccc tgtgacccct ccccagtgcc
     2581 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     2641 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     2701 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     2761 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     2821 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     2881 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     2941 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     3001 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg
     3061 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3121 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     3181 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     3241 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     3301 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     3361 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3421 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3481 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3541 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3601 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     3661 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3721 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3781 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3841 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3901 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     3961 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4021 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4081 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4141 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4201 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4261 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4321 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4381 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4441 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4501 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4561 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4621 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4681 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4741 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4801 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4861 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4921 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4981 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5041 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5101 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5161 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5221 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5281 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5341 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5401 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5461 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5521 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5581 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5641 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5701 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5761 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
