LOCUS       Exported                6404 bp ds-DNA     circular SYN 06-SEP-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6404)
  AUTHORS   HMS Neurobiology
  TITLE     Direct Submission
  JOURNAL   Exported Sep 6, 2018 from SnapGene 4.2.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6404
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    39..49
                     /label=Deleted
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     protein_bind    1450..1483
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    1534..1567
                     /label=FRT3 (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="minimal FRT site with mutations in the spacer 
                     sequence, supports FLP-mediated excision but not 
                     integration, incompatible with the WT FRT site (Schlake and
                     Bode, 1994)"
     CDS             complement(1608..2357)
                     /codon_start=1
                     /product="modified soybean ascorbate peroxidase (W41F and 
                     A134P), dimeric"
                     /label=dAPEX2
                     /translation="MGKSYPTVSADYQKAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAG
                     TFDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVA
                     VEVTGGPEVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGH
                     TIGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVD
                     KYAADEDAFFADYAEAHQKLSELGFADA"
     misc_signal     complement(2354..2363)
                     /label=Kozak consensus sequence
                     /note="vertebrate translation initiation sequence"
     protein_bind    complement(2389..2422)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(2473..2506)
                     /label=FRT3 (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="minimal FRT site with mutations in the spacer 
                     sequence, supports FLP-mediated excision but not 
                     integration, incompatible with the WT FRT site (Schlake and
                     Bode, 1994)"
     misc_feature    2531..3119
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3002..3013)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3151..3627
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   3667..3807
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3882..4337
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4619..4723
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4724..5584
                     /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"
     rep_origin      5755..6343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc
     1441 gactccggag aagttcctat tctctagaaa gtataggaac ttcgcagaat ggtagctgga
     1501 ttgtagctgc tattagcaat atgaaacctc ttggaagttc ctattcttca aatagtatag
     1561 gaacttcgcc tcaaagccgc gtgacgagct agcccatggg aattctaggc atcagcaaac
     1621 ccaagctcgg aaagcttttg gtgagcctca gcgtaatcag caaagaaggc atcttcgtcc
     1681 gctgcatatt tatcaacgag agggcggaat acagggtcag acaaaagagc cttgtcagaa
     1741 ggtagctgaa ggagaccttc cttctcacca ctcaacaact ccgtgaagta tgagttgtcg
     1801 aaaataagag gattagaggt ccagggaccc tcaaatccag aacgctcctt gtgtgcagct
     1861 ccaatagtgt gacccccaga tagagcaacg atatcttggt cagtaagccc catagctttg
     1921 ccaaacacat ctctcaaatg gtcagaaccc ttagtgggat cgggcaagcg accctctggt
     1981 ggtggctcag gcttgtcctc tcttccaggg tggaatggaa cttcaggtcc acccgtgacc
     2041 tcaacggcaa caacgccagc caactggtag aaatcggcgt agctcaaaat agggaactcc
     2101 gccttgagtg gctccaaaag cctaacagcg atgtcaagac cgttgttagc gctgtgagcc
     2161 agttcggcag ggtgcttgat ggttccgaag ggtccaccgg tcttcgtgcc cttgtcaaag
     2221 gttccagcag agtggaatgc caaacggagc attagaggag cgcatctctt ctcagcgatg
     2281 aagcctctga gcttcttctt cgccttctca acggccttct ggtaatcagc actcacagtt
     2341 gggtaagact ttcccatggt ggcaattcgt gagcgtcggt tgaacagcga agttcctata
     2401 ctttctagag aataggaact tcttgcctta acccagaaat tatcactgtt attctttaga
     2461 atggtgcaaa gagaagttcc tatactattt gaagaatagg aacttcaatc ccgatatcaa
     2521 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2581 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2641 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2701 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2761 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2821 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2881 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     2941 ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3001 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3061 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     3121 tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg
     3181 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     3241 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     3301 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     3361 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     3421 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     3481 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     3541 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     3601 tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc
     3661 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3721 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3781 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     3841 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     3901 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3961 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4021 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4081 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4141 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4201 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc
     4261 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4321 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4381 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4441 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4501 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4561 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4621 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4681 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4741 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4801 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4861 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4921 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4981 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5041 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5101 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5161 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5221 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5281 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5341 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5401 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5461 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5521 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5581 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5641 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5701 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5761 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5821 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5881 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     5941 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6001 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6061 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6121 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6181 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6241 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6301 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6361 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
