LOCUS       sequence-315118-        5814 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 5814)
  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 5814)
  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-mTurquoise2dC10-408LOV2-Ja(delta)-NES21-WPRE
FEATURES             Location/Qualifiers
     source          1..5814
                     /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    1943..2531
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1996..2016)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2414..2425)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2561..3037
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2760..2779)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3077..3217
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3077..3206
                     /label=AAV2 ITR
     rep_origin      3292..3747
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3379..3398)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3589..3610
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3764..3783)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3883..3905
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3943..3961)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4029..4133
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4134..4202,4203..4994)
                     /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(4352..4371)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5165..5753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5654..5673
                     /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 gccgccgggc tcgagagaat cgagaaaaac
     1501 ttcgtcatca ccgaccctag actgcccgat aatcctatca tcttcgcctc cgactcattt
     1561 ctccagctca ctgagtactc tcgggaggaa attctgggcc ggaattgccg gttcctccag
     1621 ggacctgaaa ccgatagagc cactgtccga aaaatccggg atgccatcga caatcagact
     1681 gaggtgaccg tccagctcat caattacacc aaatctggca aaaaattctg gaacctgttc
     1741 cacctccagc ctatgagaga tcagaagggc gacgtccagt actttattgg cgtccagctg
     1801 gatggaactg aacatgtgag ggacgctgca gaacgagagg gcgtgatgct catcaaaaaa
     1861 accgccgaaa acatcgatga gctgctgaag gagctggccg acctgaacct ggactaggga
     1921 tccgcggccg ctagataact gaaatcaacc tctggattac aaaatttgtg aaagattgac
     1981 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2041 gtatcatgct attgcttccc gtatggcctt cattttctcc tccttgtata aatcctggtt
     2101 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2161 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2221 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2281 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     2341 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     2401 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2461 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2521 cgcctccccg catcgattgt accgcgggcc cgggatctac gggtggcatc cctgtgaccc
     2581 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
     2641 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
     2701 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     2761 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     2821 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
     2881 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
     2941 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
     3001 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg
     3061 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3121 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3181 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     3241 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     3301 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3361 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3421 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3481 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3541 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3601 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     3661 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3721 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     3781 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3841 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3901 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3961 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4021 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4081 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4141 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4201 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4261 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4321 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4381 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4441 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4501 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4561 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4621 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4681 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4741 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4801 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4861 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4921 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4981 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5041 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5101 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5161 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5221 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5281 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5341 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5401 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5461 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5521 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5581 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5641 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5701 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5761 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
