LOCUS       Exported                5703 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  optoNES construct with modified mTurquoise2-LOV2 RET.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-Synapsin-NLS-mTurquoise2dC11-408LOV2-Ja(delta)-NES21-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5703)
  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 5703)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5703
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      643..652
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(730..751)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(991..1010)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1338..1359
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1832..2420
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1885..1905)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2303..2314)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2450..2926
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2649..2668)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2966..3106
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2966..3095
                     /label=AAV2 ITR
     rep_origin      3181..3636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3268..3287)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3478..3499
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3653..3672)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3772..3794
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3832..3850)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3918..4022
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4023..4883
                     /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(4241..4260)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5054..5642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5543..5562
                     /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 agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtacagctag ccgccaccat ggccgcagca
      661 aagcgcgtga agctggacct accggtcgtg agcaagggcg aggagctgtt caccggggtg
      721 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      781 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      841 aagctgcccg tgccctggcc caccctcgtg accaccctgt cctggggcgt gcagtgcttc
      901 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      961 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1021 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1081 gaggacggca acatcctggg gcacaagctg gagtacaact actttagcga caacgtctat
     1141 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     1201 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1261 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagcttag caaagacccc
     1321 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccct cgagagaatc
     1381 gagaaaaact tcgtcatcac cgaccctaga ctgcccgata atcctatcat cttcgcctcc
     1441 gactcatttc tccagctcac tgagtactct cgggaggaaa ttctgggccg gaattgccgg
     1501 ttcctccagg gacctgaaac cgatagagcc actgtccgaa aaatccggga tgccatcgac
     1561 aatcagactg aggtgaccgt ccagctcatc aattacacca aatctggcaa aaaattctgg
     1621 aacctgttcc acctccagcc tatgagagat cagaagggcg acgtccagta ctttattggc
     1681 gtccagctgg atggaactga acatgtgagg gacgctgcag aacgagaggg cgtgatgctc
     1741 atcaaaaaaa ccgccgaaaa catcgatgag ctgctgaagg agctggccga cctgaacctg
     1801 gactagggat ccgcggccgc tagataactg aaatcaacct ctggattaca aaatttgtga
     1861 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     1921 aatgcctttg tatcatgcta ttgcttcccg tatggccttc attttctcct ccttgtataa
     1981 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2041 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2101 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2161 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2221 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg
     2281 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     2341 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     2401 cctttgggcc gcctccccgc atcgattgta ccgcgggccc gggatctacg ggtggcatcc
     2461 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     2521 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     2581 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     2641 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     2701 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     2761 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     2821 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     2881 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     2941 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3001 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3061 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     3121 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     3181 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3241 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3301 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3361 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     3421 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3481 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     3541 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     3601 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     3661 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     3721 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3781 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     3841 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     3901 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3961 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4021 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4081 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4141 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4201 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4261 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4321 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4381 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4441 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4501 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4561 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4621 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4681 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4741 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4801 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4861 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4921 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4981 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5041 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5101 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5161 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5221 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5281 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5341 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5401 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5461 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5521 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5581 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5641 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5701 tgt
//
