LOCUS       sequence-315090-        5823 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 5823)
  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 5823)
  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-mTurquoise2dC7-408LOV2-Ja(delta)-NES21-WPRE
FEATURES             Location/Qualifiers
     source          1..5823
                     /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"
     regulatory      787..796
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(793..795,796..798,799..1311)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(838..859)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1099..1118)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1446..1467
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1952..2540
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2005..2025)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2423..2434)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2570..3046
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2769..2788)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3086..3226
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3086..3215
                     /label=AAV2 ITR
     rep_origin      3301..3756
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3388..3407)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3598..3619
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3773..3792)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3892..3914
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3952..3970)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4038..4142
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4143..4211,4212..5003)
                     /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(4361..4380)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5174..5762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5663..5682
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      841 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      901 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      961 ccctggccca ccctcgtgac caccctgtcc tggggcgtgc agtgcttcgc ccgctacccc
     1021 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1081 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1141 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1201 atcctggggc acaagctgga gtacaactac tttagcgaca acgtctatat caccgccgac
     1261 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     1321 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1381 cccgacaacc actacctgag cacccagtcc aagcttagca aagaccccaa cgagaagcgc
     1441 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcct cgagagaatc
     1501 gagaaaaact tcgtcatcac cgaccctaga ctgcccgata atcctatcat cttcgcctcc
     1561 gactcatttc tccagctcac tgagtactct cgggaggaaa ttctgggccg gaattgccgg
     1621 ttcctccagg gacctgaaac cgatagagcc actgtccgaa aaatccggga tgccatcgac
     1681 aatcagactg aggtgaccgt ccagctcatc aattacacca aatctggcaa aaaattctgg
     1741 aacctgttcc acctccagcc tatgagagat cagaagggcg acgtccagta ctttattggc
     1801 gtccagctgg atggaactga acatgtgagg gacgctgcag aacgagaggg cgtgatgctc
     1861 atcaaaaaaa ccgccgaaaa catcgatgag ctgctgaagg agctggccga cctgaacctg
     1921 gactagggat ccgcggccgc tagataactg aaatcaacct ctggattaca aaatttgtga
     1981 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2041 aatgcctttg tatcatgcta ttgcttcccg tatggccttc attttctcct ccttgtataa
     2101 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2161 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2221 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2281 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2341 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg
     2401 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     2461 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     2521 cctttgggcc gcctccccgc atcgattgta ccgcgggccc gggatctacg ggtggcatcc
     2581 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     2641 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     2701 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     2761 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     2821 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     2881 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     2941 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     3001 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     3061 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3121 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3181 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     3241 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     3301 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3361 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3421 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3481 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     3541 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3601 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     3661 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     3721 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     3781 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     3841 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3901 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     3961 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4021 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4081 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4141 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4201 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4261 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4321 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4381 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4441 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4501 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4561 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4621 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4681 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4741 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4801 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4861 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4921 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4981 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5041 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5101 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5161 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5221 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5281 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5341 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5401 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5461 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5521 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5581 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5641 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5701 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5761 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5821 tgt
//
