LOCUS       V020833                 5509 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020833
VERSION     V020833
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             832..855
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             859..1896
                     /label="Hmg20a(NM_025812)"
                     /note="Hmg20a(NM_025812)"
                     /gene="Hmg20a"
     CDS             1918..2637
                     /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"
     polyA_signal    2648..2782
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(2893..2909)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    2917..2933
                     /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(2941..2971)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    2986..3007
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(3295..3883)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(4054..4914)
                     /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(4915..5019)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     5493..5509
                     /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 ggcccaccat ggactacaag
      841 gacgacgatg acaagcttat ggagagcttg atggccagtt ctaccctgcc gcccctgttt
      901 gcagatgaag atggctccaa ggagagcaat gatctggcta catctgggtt aacacacccc
      961 gagggtccgt atggcagtgc agccacatca acaaccaacc cagagtttgt ggaggatctc
     1021 tcccaaggcc agctgcttca gagtgaggct tccaatgctg tagaaggcaa tgagcaaagg
     1081 cctgaagatg agcaaagaag taaacgagga ggttggtcca aagggagaaa aaggaagaaa
     1141 cctcttcgag acagcaatgc acctaaatcc ccccttacag gatatgttcg attcatgaac
     1201 gagcgtagag aacagcttcg agcaaagaga ccagaggttc catttccaga aatcacaagg
     1261 atgttgggca atgagtggag taaactgcct cctgaggaaa agcagcgcta ccttgatgaa
     1321 gcagacagag ataaggagcg ttacatgaag gaactggagc agtatcagaa gaccgaggcc
     1381 tacaaggtct tcagtaggaa aacacaggac cgtcagaaag gcaaatctca taggcaagat
     1441 gcagcccgac aggccactca tgatcatgag aaagaaacag aggtaaagga acgctctgtt
     1501 tttgatatcc ctatatttac cgaagaattc ctgaaccaca gcaaagctcg tgaggcagag
     1561 ctccgccagc ttcgtaaatc caacatggag tttgaagaga ggaacgcagc cctacagaag
     1621 cacgtggaga gcatgcgcac agcagtggag aagctggagg ttgacgtgat ccaggagcgg
     1681 agccgaaaca ccgtccttca gcagcacctg gagaccctgc ggcagatgtt gaccagcagc
     1741 ttcgccagca tgcccttgcc tggaagtgga gaaataccaa cagtggacac cattgactca
     1801 tacatgaaca gattgcacag tataatttta gctaatcccc aagacaacga aaacttcata
     1861 gctacagtcc gagaggttgt gaacaggctt gatcgtggta ccggaggagg gggaagcatg
     1921 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1981 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2041 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2101 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     2161 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2221 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     2281 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2341 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2401 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2461 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2521 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2581 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtagcgg
     2641 ccgcggggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     2701 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2761 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     2821 gggggaggtg tgggaggttt tttcggatcc tctagagtcg atctgcaggc atgctagctt
     2881 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     2941 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3001 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3061 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3121 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3181 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3241 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3301 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3361 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3421 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3481 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3541 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3601 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3661 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3721 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3781 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3841 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3901 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3961 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4021 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4081 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4141 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4201 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4261 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4321 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4381 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4441 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4501 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4561 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4621 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4681 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4741 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4801 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4861 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4921 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4981 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5041 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     5101 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     5161 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     5221 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     5281 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     5341 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     5401 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     5461 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagt
//
