LOCUS       Exported                5403 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  nucleus marker.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-Synapsin-Ypet-3XNLS-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5403)
  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 5403)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5403
                     /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      652..661
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             658..1374
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLLCTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLL
                     EFLTAAGITEGMNELYK"
     primer_bind     complement(703..724)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(964..983)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1417..1437
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    1456..1512
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    1532..2120
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1585..1605)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2003..2014)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2150..2626
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2349..2368)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2666..2806
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2666..2795
                     /label=AAV2 ITR
     rep_origin      2881..3336
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2968..2987)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3178..3199
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3353..3372)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3472..3494
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3532..3550)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3618..3722
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3723..4583
                     /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(3941..3960)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4754..5342
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5243..5262
                     /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 cgctaccggt cgccaccatg
      661 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      721 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      781 aagctgaccc tgaagcttct atgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      841 gtgaccaccc tgggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag
      901 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      961 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1021 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1081 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
     1141 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
     1201 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1261 ctgagctacc agtccgccct gttcaaagac cccaacgaga agcgcgatca catggtcctg
     1321 ctggagttcc tgaccgccgc cgggatcact gagggcatga acgagctgta caagtccgga
     1381 ctcagatccg accccagaag aaagcgcaag gtggacccca agaagaagcg caaggtggac
     1441 cccaagaaga agcgcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg
     1501 ggcccgggat ccgcggccgc tagataactg aaatcaacct ctggattaca aaatttgtga
     1561 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     1621 aatgcctttg tatcatgcta ttgcttcccg tatggccttc attttctcct ccttgtataa
     1681 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     1741 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     1801 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     1861 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     1921 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg
     1981 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     2041 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     2101 cctttgggcc gcctccccgc atcgattgta ccgcgggccc gggatctacg ggtggcatcc
     2161 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     2221 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     2281 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     2341 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     2401 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     2461 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     2521 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     2581 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     2641 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     2701 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     2761 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     2821 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     2881 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2941 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3001 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3061 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     3121 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3181 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     3241 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     3301 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     3361 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     3421 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3481 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     3541 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     3601 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3661 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3721 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3781 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3841 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3901 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3961 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4021 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4081 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4141 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4201 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4261 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4321 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4381 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4441 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4501 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4561 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4621 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4681 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4741 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4801 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4861 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4921 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4981 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5041 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5101 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5161 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5221 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5281 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5341 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5401 tgt
//
