LOCUS       V025143                 5525 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025143
VERSION     V025143
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        27..330
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        331..534
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     intron          672..768
                     /note="modified SV40 intron with splice donor and acceptor
                     sites"
                     /label="SV40 intron"
     CDS             859..1860
                     /label="Ctsl(NM_009984)"
                     /note="Ctsl(NM_009984)"
                     /gene="Ctsl"
     CDS             1897..2613
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             2623..2649
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    2664..2798
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(2909..2925)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    2933..2949
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(2957..2987)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    3002..3023
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(3311..3899)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(4070..4930)
                     /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"
     promoter        complement(4931..5035)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     5509..5525
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 gagttcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
      601 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag
      661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaggcc cgaattcggt
      841 cgaccgagat ctgccaccat gaatctttta ctccttttgg ctgtcctctg cttgggaaca
      901 gccttagcta ctccaaaatt tgatcaaacc tttagtgcag agtggcacca gtggaagtcc
      961 acacacagaa gactgtatgg cacgaatgag gaagagtgga ggagagcgat atgggagaag
     1021 aacatgagaa tgatccagct acacaacggg gaatacagca acgggcagca cggcttttcc
     1081 atggagatga acgccttcgg tgacatgacc aatgaggaat tcaggcaggt ggtgaatggc
     1141 taccgccacc agaagcacaa gaaggggagg ctttttcagg aaccgctgat gcttaagatc
     1201 cccaagtctg tggactggag agaaaagggt tgtgtgactc ctgtgaagaa ccagggccag
     1261 tgcgggtctt gttgggcgtt tagcgcatcg ggttgcctag aaggacagat gttccttaag
     1321 accggcaaac tgatctcact gagtgaacag aaccttgtgg actgttctca cgctcaaggc
     1381 aatcagggct gtaacggagg cctgatggat tttgctttcc agtacattaa ggaaaatgga
     1441 ggtctggact cggaggagtc ttacccctat gaagcgaagg acggatcttg taaatacaga
     1501 gccgagttcg ctgtggctaa tgacacaggg ttcgtggata tccctcagca agagaaagcc
     1561 ctcatgaagg ctgtggcgac tgtggggcct atttctgttg ctatggacgc aagccatccg
     1621 tctctccagt tctatagttc aggcatctac tatgaaccca actgtagcag caagaacctc
     1681 gaccatgggg ttctgttggt gggctatggc tatgaaggaa cagattcaaa taagaataaa
     1741 tattggcttg tcaagaacag ctggggaagt gaatggggta tggaaggcta catcaaaata
     1801 gccaaagacc gggacaacca ctgtggactt gccaccgcgg ccagctatcc tgtcgtgaat
     1861 ctcgagggtg gcggtggatc aggaggaggt ggctctatgg tgagcaaggg cgaggagctg
     1921 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1981 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     2041 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     2101 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2161 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2221 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2281 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     2341 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     2401 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     2461 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     2521 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2581 gggatcactc tcggcatgga cgagctgtac aagggtaccg gctacccata cgatgttcca
     2641 gattacgctt aagcggccgc ggggatccag acatgataag atacattgat gagtttggac
     2701 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     2761 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     2821 ttatgtttca ggttcagggg gaggtgtggg aggttttttc ggatcctcta gagtcgatct
     2881 gcaggcatgc tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2941 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3001 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3061 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3121 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3181 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3241 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3301 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3361 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3421 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3481 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3541 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3601 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3661 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3721 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     3781 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3841 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3901 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3961 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4021 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4081 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4141 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4201 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4261 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4321 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4381 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4441 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4501 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4561 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4621 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4681 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4741 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4801 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4861 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4921 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4981 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5041 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     5101 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     5161 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     5221 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     5281 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     5341 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     5401 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     5461 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     5521 ccagt
//
