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 //