LOCUS       V028966                 7900 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028966
VERSION     V028966
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             874..1590
                     /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             1627..3582
                     /label="CD93(NM_012072)"
                     /note="CD93(NM_012072)"
                     /gene="CD93"
     polyA_signal    3662..3886
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3932..4360
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4374..4703
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4770..5369
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    5543..5664
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5713..5729)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5737..5753
                     /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(5761..5791)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5806..5827
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6115..6700)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6871..7731)
                     /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(7732..7836)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag aagcttggtg gaggcggatc aggaggcggt
     1621 ggatctatgg ccacctccat gggcctgctg ctgctgctgc tgctgctcct gacccagccc
     1681 ggggcgggga cgggagctga cacggaggcg gtggtctgcg tggggaccgc ctgctacacg
     1741 gcccactcgg gcaagctgag cgctgccgag gcccagaacc actgcaacca gaacgggggc
     1801 aacctggcca ctgtgaagag caaggaggag gcccagcacg tccagcgagt actggcccag
     1861 ctcctgaggc gggaggcagc cctgacggcg aggatgagca agttctggat tgggctccag
     1921 cgagagaagg gcaagtgcct ggaccctagt ctgccgctga agggcttcag ctgggtgggc
     1981 gggggggagg acacgcctta ctctaactgg cacaaggagc tccggaactc gtgcatctcc
     2041 aagcgctgtg tgtctctgct gctggacctg tcccagccgc tccttcccag ccgcctcccc
     2101 aagtggtctg agggcccctg tgggagccca ggctcccccg gaagtaacat tgagggcttc
     2161 gtgtgcaagt tcagcttcaa aggcatgtgc cggcctctgg ccctgggggg cccaggtcag
     2221 gtgacctaca ccaccccctt ccagaccacc agttcctcct tggaggctgt gccctttgcc
     2281 tctgcggcca atgtagcctg tggggaaggt gacaaggacg agactcagag tcattatttc
     2341 ctgtgcaagg agaaggcccc cgatgtgttc gactggggca gctcgggccc cctctgtgtc
     2401 agccccaagt atggctgcaa cttcaacaat gggggctgcc accaggactg ctttgaaggg
     2461 ggggatggct ccttcctctg cggctgccga ccaggattcc ggctgctgga tgacctggtg
     2521 acctgtgcct ctcgaaaccc ttgcagctcc agcccatgtc gtgggggggc cacgtgcgtc
     2581 ctgggacccc atgggaaaaa ctacacgtgc cgctgccccc aagggtacca gctggactcg
     2641 agtcagctgg actgtgtgga cgtggatgaa tgccaggact ccccctgtgc ccaggagtgt
     2701 gtcaacaccc ctgggggctt ccgctgcgaa tgctgggttg gctatgagcc gggcggtcct
     2761 ggagaggggg cctgtcagga tgtggatgag tgtgctctgg gtcgctcgcc ttgcgcccag
     2821 ggctgcacca acacagatgg ctcatttcac tgctcctgtg aggagggcta cgtcctggcc
     2881 ggggaggacg ggactcagtg ccaggacgtg gatgagtgtg tgggcccggg gggccccctc
     2941 tgcgacagct tgtgcttcaa cacacaaggg tccttccact gtggctgcct gccaggctgg
     3001 gtgctggccc caaatggggt ctcttgcacc atggggcctg tgtctctggg accaccatct
     3061 gggccccccg atgaggagga caaaggagag aaagaaggga gcaccgtgcc ccgtgctgca
     3121 acagccagtc ccacaagggg ccccgagggc acccccaagg ctacacccac cacaagtaga
     3181 ccttcgctgt catctgacgc ccccatcaca tctgccccac tcaagatgct ggcccccagt
     3241 gggtccccag gcgtctggag ggagcccagc atccatcacg ccacagctgc ctctggcccc
     3301 caggagcctg caggtgggga ctcctccgtg gccacacaaa acaacgatgg cactgacggg
     3361 caaaagctgc ttttattcta catcctaggc accgtggtgg ccatcctact cctgctggcc
     3421 ctggctctgg ggctactggt ctatcgcaag cggagagcga agagggagga gaagaaggag
     3481 aagaagcccc agaatgcggc agacagttac tcctgggttc cagagcgagc tgagagcagg
     3541 gccatggaga accagtacag tccgacacct gggacagact gctgagaatt ctgcagatat
     3601 ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     3661 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3721 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3781 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3841 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3901 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3961 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4021 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4081 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4141 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4201 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4261 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4321 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4381 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4441 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4501 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4561 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4621 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4681 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4741 atcaagagac aggatgagga tcgtttcgca tgaccgagta caagcccacg gtgcgcctcg
     4801 ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc
     4861 ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag
     4921 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     4981 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     5041 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     5101 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     5161 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     5221 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     5281 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     5341 ggtgcatgac ccgcaagccc ggtgcctgag cgggactctg gggttcgaaa tgaccgacca
     5401 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5461 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5521 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5581 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5641 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5701 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5761 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5821 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5881 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5941 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6001 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6061 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6121 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6181 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6241 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6301 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6361 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6421 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6481 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6541 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6601 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     6661 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6721 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6781 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6841 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6901 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6961 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7021 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7081 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7141 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7201 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7261 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7321 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7381 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7441 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7501 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7561 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7621 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7681 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7741 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7801 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7861 tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
