LOCUS       Exported                5824 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses pAAV-hSynapsin-Flpo.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hSynapsin-Flpo
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5824)
  AUTHORS   Xue M, Atallah BV, Scanziani M
  TITLE     Equalizing excitation-inhibition ratios across visual cortical 
            neurons.
  JOURNAL   Nature. 2014 Jul 31;511(7511):596-600. doi: 10.1038/nature13321. 
            Epub 2014 Jun 22.
  PUBMED    25043046
REFERENCE   2  (bases 1 to 5824)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5824
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   2..131
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     polyA_signal    187..235
                     /note="synthetic polyadenylation signal"
     misc_feature    249..340
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     289..308
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        376..823
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      854..863
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             860..2158
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     misc_feature    2177..2765
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2230..2250)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2648..2659)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2798..2815)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2804..3011
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3020..3149
                     /label=AAV2 ITR
     rep_origin      3253..3708
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3340..3359)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3550..3571
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3775..3794)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3894..3916
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3954..3972)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4040..4144
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4145..5005
                     /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(4363..4382)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5176..5764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5665..5684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
       61 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      121 taggggttcc tgcggccgca cgcgttctag aaatattaag gtacgggagg tacttggagc
      181 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
      241 atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag
      301 gctgtcccca gtgcaagtgc aggtgccaga acatttctct attctagact cgagctgcag
      361 agggccctgc gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc
      421 ctacctgacg accgaccccg acccactgga caagcaccca acccccattc cccaaattgc
      481 gcatccccta tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc
      541 agcttcagca ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct
      601 cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca
      661 ccttggtcgc gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat
      721 aggggggcac gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc
      781 tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg cagtcgaatt caagctgcta
      841 gcaaggatcc gccgccacca tggctcctaa gaagaagagg aaggtgatga gccagttcga
      901 catcctgtgc aagacccccc ccaaggtgct ggtgcggcag ttcgtggaga gattcgagag
      961 gcccagcggc gagaagatcg ccagctgtgc cgccgagctg acctacctgt gctggatgat
     1021 cacccacaac ggcaccgcca tcaagagggc caccttcatg agctacaaca ccatcatcag
     1081 caacagcctg agcttcgaca tcgtgaacaa gagcctgcag ttcaagtaca agacccagaa
     1141 ggccaccatc ctggaggcca gcctgaagaa gctgatcccc gcctgggagt tcaccatcat
     1201 cccttacaac ggccagaagc accagagcga catcaccgac atcgtgtcca gcctgcagct
     1261 gcagttcgag agcagcgagg aggccgacaa gggcaacagc cacagcaaga agatgctgaa
     1321 ggccctgctg tccgagggcg agagcatctg ggagatcacc gagaagatcc tgaacagctt
     1381 cgagtacacc agcaggttca ccaagaccaa gaccctgtac cagttcctgt tcctggccac
     1441 attcatcaac tgcggcaggt tcagcgacat caagaacgtg gaccccaaga gcttcaagct
     1501 ggtgcagaac aagtacctgg gcgtgatcat tcagtgcctg gtgaccgaga ccaagacaag
     1561 cgtgtccagg cacatctact ttttcagcgc cagaggcagg atcgaccccc tggtgtacct
     1621 ggacgagttc ctgaggaaca gcgagcccgt gctgaagaga gtgaacagga ccggcaacag
     1681 cagcagcaac aagcaggagt accagctgct gaaggacaac ctggtgcgca gctacaacaa
     1741 ggccctgaag aagaacgccc cctaccccat cttcgctatc aagaacggcc ctaagagcca
     1801 catcggcagg cacctgatga ccagctttct gagcatgaag ggcctgaccg agctgacaaa
     1861 cgtggtgggc aactggagcg acaagagggc ctccgccgtg gccaggacca cctacaccca
     1921 ccagatcacc gccatccccg accactactt cgccctggtg tccaggtact acgcctacga
     1981 ccccatcagc aaggagatga tcgccctgaa ggacgagacc aaccccatcg aggagtggca
     2041 gcacatcgag cagctgaagg gcagcgccga gggcagcatc agataccccg cctggaacgg
     2101 catcatcagc caggaggtgc tggactacct gagcagctac atcaacaggc ggatctgaga
     2161 attcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2221 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2281 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     2341 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     2401 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     2461 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2521 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     2581 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgc ccttctgcta
     2641 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     2701 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     2761 cccgcatcga taccgtcgac tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     2821 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2881 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2941 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3001 catgctgggg agagatctga ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     3061 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     3121 gcggcctcag tgagcgagcg agcgcgcaga gagggagtgg cccccccccc cccccctgca
     3181 gcctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     3241 aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3301 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3361 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3421 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3481 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3541 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3601 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3661 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     3721 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
     3781 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3841 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3901 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3961 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4021 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4081 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4141 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4201 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4261 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4321 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4381 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4441 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4501 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4561 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4621 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4681 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4741 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4801 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4861 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4921 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4981 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5041 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5101 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5161 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5221 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5281 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5341 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5401 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5461 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5521 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5581 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5641 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5701 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5761 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5821 catg
//
