LOCUS       62056_2440        6141 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6141)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     enhancer        221..524
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        525..727
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          876..1254
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and
                     human beta-globin intron 2"
     promoter        1325..1343
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             1421..1492
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags"
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     misc_feature    1528..2111
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2112..2828
                     /codon_start=1
                     /label=hrGFP
                     /note="humanized Renilla green fluorescent protein"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     promoter        complement(2862..2880)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2888..3364
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   3404..3544
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3619..4074
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4356..4460
                     /label=AmpR promoter
     CDS             4461..5318
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5492..6080
                     /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 cgcgtggagc tagttattaa tagtaatcaa
      181 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      241 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      301 ttcccatagt aacgtcaata gggactttcc attgacgtca atgggtggag tatttacggt
      361 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      421 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      481 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      541 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      601 ttgacgtcaa tgggagtttg ttttgcacca aaatcaacgg gactttccaa aatgtcgtaa
      661 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      721 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
      781 ccatagaaga caccgggacc gatccagcct ccgcggattc gaatcccggc cgggaacggt
      841 gcattggaac gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata
      901 ggcccacaaa aaatgctttc ttcttttaat atactttttt gtttatctta tttctaatac
      961 tttccctaat ctctttcttt cagggcaata atgatacaat gtatcatgcc tctttgcacc
     1021 attctaaaga ataacagtga taatttctgg gttaaggcaa tagcaatatt tctgcatata
     1081 aatatttctg catataaatt gtaactgatg taagaggttt catattgcta atagcagcta
     1141 caatccagct accattctgc ttttatttta tggttgggat aaggctggat tattctgagt
     1201 ccaagctagg cccttttgct aatcatgttc atacctctta tcttcctccc acagctcctg
     1261 ggcaacgtgc tggtctgtgt gctggcccat cactttggca aagaattggg attcgaacat
     1321 cgataattaa ccctcactaa agggaacaaa agctggagct ccaccgcggt ggcggccgct
     1381 ctagcccggg cggatccgaa ttcgcatgcg tcgactcgag gactacaagg atgacgatga
     1441 caaggattac aaagacgacg atgataagga ctataaggat gatgacgaca aataatagca
     1501 attcctcgac gactgcatag ggttaccccc ctctccctcc ccccccccta acgttactgg
     1561 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
     1621 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     1681 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     1741 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     1801 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
     1861 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
     1921 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
     1981 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa
     2041 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     2101 tggccacaac catggtgagc aagcagatcc tgaagaacac cggcctgcag gagatcatga
     2161 gcttcaaggt gaacctggag ggcgtggtga acaaccacgt gttcaccatg gagggctgcg
     2221 gcaagggcaa catcctgttc ggcaaccagc tggtgcagat ccgcgtgacc aagggcgccc
     2281 ccctgccctt cgccttcgac atcctgagcc ccgccttcca gtacggcaac cgcaccttca
     2341 ccaagtaccc cgaggacatc agcgacttct tcatccagag cttccccgcc ggcttcgtgt
     2401 acgagcgcac cctgcgctac gaggacggcg gcctggtgga gatccgcagc gacatcaacc
     2461 tgatcgagga gatgttcgtg taccgcgtgg agtacaaggg ccgcaacttc cccaacgacg
     2521 gccccgtgat gaagaagacc atcaccggcc tgcagcccag cttcgaggtg gtgtacatga
     2581 acgacggcgt gctggtgggc caggtgatcc tggtgtaccg cctgaacagc ggcaagttct
     2641 acagctgcca catgcgcacc ctgatgaaga gcaagggcgt ggtgaaggac ttccccgagt
     2701 accacttcat ccagcaccgc ctggagaaga cctacgtgga ggacggcggc ttcgtggagc
     2761 agcacgagac cgccatcgcc cagctgacca gcctgggcaa gcccctgggc agcctgcacg
     2821 agtgggtgta atagggtacc aggtaagtgt acccaattcg ccctatagtg agtcgtatta
     2881 gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     2941 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     3001 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     3061 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca
     3121 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc
     3181 cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta
     3241 gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta
     3301 cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt
     3361 ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     3421 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     3481 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     3541 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3601 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3661 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3721 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3781 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     3841 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3901 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3961 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4021 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     4081 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4141 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4201 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4261 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4321 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4381 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4441 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4501 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4561 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4621 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4681 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4741 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4801 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4861 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4921 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4981 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5041 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5101 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5161 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5221 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5281 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5341 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5401 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5461 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5521 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5581 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     5641 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5701 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5761 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5821 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5881 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5941 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6001 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6061 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6121 gctggccttt tgctcacatg t
//
