LOCUS       V044213                 8813 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044213
VERSION     V044213
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             962..1675
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="enhanced YFP"
                     /transl_table=1
                     /translation="V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
                     /label="EYFP"
     CDS             1715..5143
                     /label="MCM9(NM_017696)"
                     /note="MCM9(NM_017696)"
                     /gene="MCM9"
     polyA_signal    5154..5776
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        5805..6134
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(6173..6191)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(6205..6221)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6229..6245
                     /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(6253..6283)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6298..6319
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6607..7195)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7366..8226)
                     /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(8227..8331)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      8358..8786
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
      961 tgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1021 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1081 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1141 cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca
     1201 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1261 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1321 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1381 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1441 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1501 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1561 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1621 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaaggcggc
     1681 cgcgggtgga ggcggatcag gaggcggtgg atctatgaat agcgatcaag ttacactggt
     1741 tggtcaagtg tttgagtcat atgtttcgga ataccataag aatgatattc ttctaatctt
     1801 gaaggaaagg gatgaagatg ctcattaccc agttgtggtt aatgccatga ctctgtttga
     1861 gaccaacatg gaaatcgggg aatatttcaa catgttcccc agtgaagtgc ttacaatttt
     1921 tgatagtgca ctgcgaaggt cagccttgac aattctccag tccctttctc agcctgaggc
     1981 tgtttccatg aaacagaatc ttcatgccag gatatcaggt ttgcctgtct gtcctgagct
     2041 ggtgagggaa cacataccta aaaccaagga tgtgggacac tttttatctg tcactgggac
     2101 agtgattcga acaagtctgg tgaaggttct ggagtttgag cgggattaca tgtgtaacaa
     2161 atgcaagcat gtgtttgtga tcaaggctga ctttgagcag tattacacct tttgccggcc
     2221 atcctcgtgt cccagcttgg agagctgtga ttcctctaaa ttcacttgcc tctcaggctt
     2281 gtcttcgtct ccaaccaggt gtagagatta ccaggaaatc aaaattcagg aacaggttca
     2341 aaggctatct gttggaagta ttccacgatc tatgaaggtt attctggaag atgacttagt
     2401 ggatagttgc aaatctggtg atgacctcac tatttacggg attgtaatgc aacggtggaa
     2461 gccctttcag caagatgtgc gctgtgaagt ggagatagtc ctgaaagcaa attacatcca
     2521 agtaaataat gagcagtcct cagggatcat catggatgag gaggtccaaa aggaattcga
     2581 agatttttgg gaatactata agagcgatcc ctttgcagga aggaatgtaa tattggctag
     2641 cttgtgccct caagtgtttg gaatgtatct agtaaagctt gctgtggcca tggtgctggc
     2701 tggtgggatt caaaggactg atgctacagg aacacgggtc agaggagaat ctcatctttt
     2761 attggttggg gatcctggca cagggaaatc tcagttcctc aaatatgcag caaagattac
     2821 accaagatct gtgctgacca caggaattgg atctactagt gcaggtctga cggtaactgc
     2881 tgtaaaagac tcaggagaat ggaatttgga ggctggggca ttagttcttg cagatgcggg
     2941 cctttgctgt attgatgagt tcaatagcct caaagagcat gataggacca gtatccatga
     3001 agcaatggag caacaaacca taagtgttgc taaggctggc ctcgtgtgca agctgaacac
     3061 aaggaccacc atcctggcag caacgaaccc caaaggccag tacgaccccc aggagtccgt
     3121 gtctgtgaac attgccctcg gcagcccact cttaagtcga tttgacctga tcctggtttt
     3181 gcttgatacc aagaatgaag actgggatcg tatcatttcc tcctttatct tagaaaataa
     3241 aggttaccca agcaaatcag agaagctctg gagcatggaa aagatgaaaa cctatttctg
     3301 cctcataagg aatctgcagc ccacactgtc tgatgtgggc aatcaggttc ttctccggta
     3361 ctaccagatg caaaggcaga gtgattgccg gaacgctgcc cggaccacca ttcggctgtt
     3421 ggaaagcttg atacgattag cagaagctca tgctcgcctg atgtttcgtg atactgtaac
     3481 tctggaagac gctattacgg tggtgtcagt catggagtcc tcaatgcagg gaggtgcact
     3541 gctaggaggt gtgaatgccc tccacacttc ctttcctgaa aaccctggag agcagtacca
     3601 gagacagtgt gaacttattc tggaaaagct agagctgcag agcctcttga gtgaagagct
     3661 tagaagactt gaaaggttac agaatcagag tgtgcaccaa tcccaaccac gggtattgga
     3721 ggtagagact actccaggat ccttgagaaa tggtccaggg gaagaatcaa acttcagaac
     3781 ttcatcacag caggaaatca actatagcac acatatcttc tctcctggag gcagccccga
     3841 gggaagccca gttctagatc ccccaccgca tctggagcct aatagatcaa caagtaggaa
     3901 acattcagct cagcacaaaa ataacagaga tgacagttta gattggtttg atttcatggc
     3961 aactcatcag agtgaaccta aaaacactgt tgttgtgtct cctcatccca aaacatctgg
     4021 agaaaatatg gcttcgaaga tctctaacag cacatctcag ggtaaggaga agagtgagcc
     4081 aggccaaagg agcaaagtgg acattgggtt gcttccatca ccaggagaga caggtgttcc
     4141 atggagggca gacaatgtgg aaagtaacaa gaaaaaaagg ctagcactag attctgaagc
     4201 agcagtctct gctgataaac cagactcagt actgactcat catgtcccca ggaacctgca
     4261 gaagctgtgc aaagagaggg cccagaagtt gtgcagaaat agcaccaggg tgcctgcaca
     4321 gtgcacagtc ccttcccatc ctcagtccac tcctgtacat agcccagaca gaatgctgga
     4381 ctcacccaaa agaaagagac cgaaatccct tgcgcaagtg gaagagcctg caattgaaaa
     4441 tgttaagcct ccaggttccc ctgtggccaa actggcaaaa tttactttca agcagaagtc
     4501 aaaactgatc cactcctttg aagatcacag ccatgtgtca cctggtgcaa ctaaaatagc
     4561 agttcatagt cctaaaattt cccagcgtag aacaagaaga gacgcagcct tgccggtgaa
     4621 gcgtccagga aagttaacat ctaccccagg aaaccagatc tccagtcagc cacagggtga
     4681 gacaaaggag gtgtcgcagc agccaccaga gaaacacgga ccaagagaga aggtgatgtg
     4741 tgcccctgag aagaggatta ttcagcctga attagagctt gggaacgaga ctgggtgtgc
     4801 tcatcttact tgtgagggag acaaaaagga agaggtttca ggcagtaata aaagcggcaa
     4861 ggttcatgcc tgcacattag ccagattggc aaacttctgc tttactcccc catcggaatc
     4921 caaatcaaaa tcccctcctc ctgaaaggaa gaaccgaggt gagagaggcc caagctcccc
     4981 tcctacaacc acagctccaa tgcgtgtcag taaaaggaaa tcttttcagc tccgtgggtc
     5041 caccgagaaa ctgattgttt ccaaagaatc cctcttcact ttaccagaac taggtgatga
     5101 agcatttgat tgtgactggg atgaagagat gagaaaaaag tcatagggat cccgggtggc
     5161 atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca
     5221 ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata
     5281 atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta
     5341 gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg
     5401 caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat
     5461 tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca
     5521 ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc
     5581 ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa
     5641 aataactata ccagcaggag gacgtccaga cacagcatag gctacctggc catgcccaac
     5701 cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg tccactcagt
     5761 agatgcctgt tgaattgggt acgcggccag cttggctgtg gaatgtgtgt cagttagggt
     5821 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5881 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5941 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     6001 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     6061 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     6121 taggcttttg caaaaagctc ctcgaggaac tgaaaaacca gaaagttaat tccctatagt
     6181 gagtcgtatt aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct
     6241 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6301 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6361 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6421 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6481 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6541 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6601 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6661 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6721 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6781 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6841 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6901 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6961 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7021 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     7081 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7141 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7201 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7261 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7321 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7381 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7441 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7501 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7561 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7621 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7681 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7741 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7801 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7861 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7921 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7981 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8041 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8101 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8161 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8221 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8281 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8341 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     8401 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     8461 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     8521 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     8581 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     8641 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     8701 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     8761 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa ttt
//
