LOCUS       V030492                 6864 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030492
VERSION     V030492
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     repeat_region   1..141
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
                     /rpt_type=inverted
                     /label="AAV2 ITR"
     enhancer        156..535
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        536..739
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             752..2128
                     /label="Itpka(NM_031045)"
                     /note="Itpka(NM_031045)"
                     /gene="Itpka"
     CDS             2129..2194
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2210..2263
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             2273..2968
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="Zoanthus green fluorescent protein"
                     /transl_table=1
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP*"
                     /label="ZsGreen1"
     misc_feature    2991..3579
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3611..4087
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     repeat_region   4127..4267
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
                     /rpt_type=inverted
                     /label="AAV2 ITR"
     rep_origin      4342..4797
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5079..5183
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             5184..6044
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      6215..6803
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgacat tgattattga ctagttatta
      181 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      241 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      301 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      361 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      421 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      481 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      541 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      601 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      661 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      721 ggtctatata agcagagctg gtaccgccac catgaccctg cccggacacc cgacgggcat
      781 ggcgcgacca cggggcgcgg ggccctgcag ccccgggttg gagcgggctc cgcgccggag
      841 cgtcggggag ctgcgcctgc tcttcgaagc gcgctgcgcc gcagtcgccg ccgcagcggc
      901 cgcaggggag ccccgggccc gcggggccaa gcggcgtggg ggacaagtgc ctaacgggct
      961 cccgcgtgct gcccctgctc cggtgatccc gcagctcact gtgacaagcg aggaggatgt
     1021 ggccccggcc agccccgggc cgccggaccg ggaggggaac tggctcccag ctgcggggtc
     1081 gcacctgcag cagccacgcc gcctctccac ctcgtccctc tcctccaccg gctcctcgtc
     1141 gctgctcgag gactcggagg acgacctgct gagcgacagc gagagccgga gccgcggcaa
     1201 cgtgcagctg gaaaccagcg aagacgtggg gcagaaaagc cattggcaga agatccgtac
     1261 catggtcaac ctgcctgtca tgagtccttt caaaaagcgc tactcctggg tgcagttagc
     1321 agggcacaca gggagtttca aagctgcggg caccagcggc ctgatcctga agcgcagctc
     1381 ggagcctgaa cactactgcc tggtgcggtt gatggctgat gtgctgcgtg ggtgtgtgcc
     1441 ggccttccac ggtgtggtcg agcgggatgg tgaaagctac ttacagttac aggacctgct
     1501 tgatggcttc gatggacctt gcgtgcttga ctgcaagatg ggtgtcagaa cttacctgga
     1561 agaagagctg accaaagccc gagaacggcc caagctgcga aaggacatgt ataagaaaat
     1621 gctggccgtg gaccccgagg cacctactga ggaggagcat gcgcagcgtg ccgtcacaaa
     1681 acctcgatac atgcaatggc gcgaaggcat cagctccagc actacactcg gcttccgcat
     1741 cgagggcatc aagaaagccg atggatcttg cagtactgat ttcaaaacta cacgaagcag
     1801 agagcaagtg actcgagtct ttgaggagtt catgcaagga gatgctgaag tgctgaagag
     1861 gtatctgaac cgcctacagc agatccggga taccctggag atctctgatt tctttagacg
     1921 gcacgaggtg attggcagct cactcctctt cgtgcatgac cattgccatc gcgctggtgt
     1981 gtggctcatc gattttggca agaccacgcc tctccccgat ggccagatcc tggatcatcg
     2041 gaggccctgg gaggagggca atcgtgagga cggctatttg ctggggctgg acaatctcat
     2101 tggcatcttg gccaacctgg ctgagagaga ctacaaagac catgacggtg attataaaga
     2161 tcatgacatc gattacaagg atgacgatga caagcctcta gagtcgacag agggcagagg
     2221 aagtcttcta acatgcggtg acgtggagga gaatcccggc ccttccggaa ccatggccca
     2281 gtccaagcac ggcctgacca aggagatgac catgaagtac cgcatggagg gctgcgtgga
     2341 cggccacaag ttcgtgatca ccggcgaggg catcggctac cccttcaagg gcaagcaggc
     2401 catcaacctg tgcgtggtgg agggcggccc cttgcccttc gccgaggaca tcttgtccgc
     2461 cgccttcatg tacggcaacc gcgtgttcac cgagtacccc caggacatcg tcgactactt
     2521 caagaactcc tgccccgccg gctacacctg ggaccgctcc ttcctgttcg aggacggcgc
     2581 cgtgtgcatc tgcaacgccg acatcaccgt gagcgtggag gagaactgca tgtaccacga
     2641 gtccaagttc tacggcgtga acttccccgc cgacggcccc gtgatgaaga agatgaccga
     2701 caactgggag ccctcctgcg agaagatcat ccccgtgccc aagcagggca tcttgaaggg
     2761 cgacgtgagc atgtacctgc tgctgaagga cggtggccgc ttgcgctgcc agttcgacac
     2821 cgtgtacaag gccaagtccg tgccccgcaa gatgcccgac tggcacttca tccagcacaa
     2881 gctgacccgc gaggaccgca gcgacgccaa gaaccagaag tggcacctga ccgagcacgc
     2941 catcgcctcc ggctccgcct tgccctgaat tcgatatcaa gcttatcgat aatcaacctc
     3001 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     3061 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     3121 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     3181 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     3241 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     3301 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     3361 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     3421 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     3481 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     3541 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg
     3601 agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
     3661 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
     3721 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
     3781 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
     3841 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc
     3901 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
     3961 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
     4021 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
     4081 tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg
     4141 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     4201 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     4261 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4321 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4381 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4441 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4501 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     4561 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4621 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4681 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4741 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     4801 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     4861 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4921 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4981 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     5041 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5101 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5161 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5221 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5281 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5341 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5401 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5461 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5521 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5581 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5641 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5701 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5761 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5821 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5881 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5941 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6001 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6061 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6121 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6181 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6241 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6301 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6361 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6421 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6481 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6541 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6601 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6661 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6721 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6781 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6841 tttgctggcc ttttgctcac atgt
//
