LOCUS       Exported                6524 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 6524)
  AUTHORS   Qiyu Zhang
  TITLE     Direct Submission
  JOURNAL   Exported Sep 6, 2018 from SnapGene 4.2.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6524
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     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..2354)
                     /codon_start=1
                     /product="modified soybean ascorbate peroxidase (W41F and 
                     A134P), dimeric"
                     /label=dAPEX2
                     /translation="GKSYPTVSADYQKAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
                     FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
                     EVTGGPEVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
                     IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
                     YAADEDAFFADYAEAHQKLSELGFADA"
     CDS             complement(2355..2396)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(2406..2477)
                     /codon_start=1
                     /label=COX4 localization signal
                     /translation="MLATRVFSLVGKRAISTSVCVRAH"
     misc_feature    complement(2478..2483)
                     /label=Kozak
     protein_bind    complement(2509..2542)
                     /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(2593..2626)
                     /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    2651..3239
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3122..3133)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3271..3747
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   3787..3927
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4002..4457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4739..4843
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4844..5704
                     /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      5875..6463
                     /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 ttccggtgct gtccaggccc agcagggggt tggggatggg cttgcctgga
     2401 tccttgtggg cgcgcacgca cacgctggtg ctgatggcgc gcttgcccac caggctgaac
     2461 acgcgggtgg ccagcatggt ggcaattcgt gagcgtcggt tgaacagcga agttcctata
     2521 ctttctagag aataggaact tcttgcctta acccagaaat tatcactgtt attctttaga
     2581 atggtgcaaa gagaagttcc tatactattt gaagaatagg aacttcaatc ccgatatcaa
     2641 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2701 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2761 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2821 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2881 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2941 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3001 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     3061 ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3121 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3181 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     3241 tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg
     3301 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     3361 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     3421 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     3481 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     3541 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     3601 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     3661 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     3721 tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc
     3781 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3841 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3901 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     3961 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     4021 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4081 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4141 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4201 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4261 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4321 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc
     4381 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4441 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4501 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4561 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4621 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4681 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4741 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4801 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4861 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4921 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4981 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5041 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5101 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5161 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5221 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5281 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5341 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5401 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5461 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5521 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5581 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5641 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5701 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5761 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5821 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5881 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5941 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6001 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     6061 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6121 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6181 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6241 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6301 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6361 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6421 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6481 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
