LOCUS       V018342                 8347 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018342
VERSION     V018342
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        236..615
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        616..819
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        864..882
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             924..3125
                     /label="MCM5(NM_006739)"
                     /note="MCM5(NM_006739)"
                     /gene="MCM5"
     CDS             3177..3896
                     /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    3948..4172
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4218..4646
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4660..4989
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5056..5850
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6024..6145
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6194..6210)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6218..6234
                     /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(6242..6272)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6287..6308
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6596..7181)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7352..8212)
                     /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(8213..8317)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
       61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
      121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      241 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      601 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      781 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
      841 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag
      901 cgtttaaact taagcttgcc accatgtcgg gattcgacga tcctggcatt ttctacagcg
      961 acagcttcgg gggcgacgcc caggccgacg aggggcaggc ccgcaaatcg cagctgcaga
     1021 ggcgcttcaa ggagttcctg cggcagtacc gagtgggcac cgaccgcacg ggcttcacct
     1081 tcaaatacag ggatgaactc aagcggcatt acaacctggg ggagtactgg attgaggtgg
     1141 agatggagga tctggccagc tttgatgagg acctggccga ctacttgtac aagcagccag
     1201 ccgagcacct gcagctgctg gaggaagctg ccaaggaggt agctgatgag gtgacccggc
     1261 cccggccttc tggggaggag gtgctccagg acatccaggt catgctcaag tcggacgcca
     1321 gcccttccag cattcgtagc ctgaagtcgg acatgatgtc acacctggtg aagatccctg
     1381 gcatcatcat cgcggcctct gcggtccgtg ccaaggccac ccgcatctct atccagtgcc
     1441 gcagctgccg caacaccctc accaacattg ccatgcgccc tggcctcgag ggctatgccc
     1501 tgcccaggaa gtgcaacaca gatcaggctg ggcgccccaa atgcccattg gacccgtact
     1561 tcatcatgcc cgacaaatgc aaatgcgtgg acttccagac cctgaagctg caggagctgc
     1621 ctgatgcagt cccccacggg gagatgccca gacacatgca gctctactgc gacaggtacc
     1681 tgtgtgacaa ggtcgtccct gggaacaggg ttaccatcat gggcatctac tccatcaaga
     1741 agtttggcct gactaccagc aggggccgtg acagggtggg cgtgggcatc cgaagctcct
     1801 acatccgtgt cctgggcatc caggtggaca cagatggctc tggccgcagc tttgctgggg
     1861 ccgtgagccc ccaggaggag gaggagttcc gtcgcctggc tgccctccca aatgtctatg
     1921 aggtcatctc caagagcatc gccccctcca tctttggggg cacagacatg aagaaggcca
     1981 ttgcctgcct gctctttggg ggctcccgaa agaggctccc tgatggactt actcgccgag
     2041 gagacatcaa cctgctgatg ctaggggacc ctgggacagc caagtcccag cttctgaagt
     2101 ttgtggagaa gtgttctccc attggggtat acacgtctgg gaaaggcagc agcgcagctg
     2161 gactgacagc ctcggtgatg agggaccctt cgtcccggaa tttcatcatg gagggcggag
     2221 ccatggtcct ggccgatggt ggggtcgtct gtattgacga gtttgacaag atgcgagaag
     2281 atgaccgtgt ggcaatccac gaagccatgg agcagcagac catctctatc gccaaggctg
     2341 ggatcaccac caccctgaac tcccgctgct ccgtcctggc tgctgccaac tcagtgttcg
     2401 gccgctggga tgagacgaag ggggaggaca acattgactt catgcccacc atcttgtcgc
     2461 gcttcgacat gatcttcatc gtcaaggatg agcacaatga ggagagggat gtgatgctgg
     2521 ccaagcatgt catcactctg cacgtgagcg cactgacaca gacacaggct gtggagggcg
     2581 agattgacct ggccaagctg aagaagttta ttgcctactg ccgagtgaag tgtggccccc
     2641 ggctgtcagc agaggctgca gagaaactga agaaccgcta catcatcatg cggagcgggg
     2701 cccgtcagca cgagagggac agtgaccgcc gctccagcat ccccatcact gtgcggcagc
     2761 tggaggccat tgtgcgcatc gcggaagccc tcagcaagat gaagctgcag cccttcgcca
     2821 cagaggcaga tgtggaggag gccctgcggc tcttccaagt gtccacgttg gatgctgcct
     2881 tgtccggtac cctgtcaggg gtggagggct tcaccagcca ggaggaccag gagatgctga
     2941 gccgcatcga gaagcagctc aagcgccgct ttgccattgg ctcccaggtg tctgagcaca
     3001 gcatcatcaa ggacttcacc aagcagaaat acccggagca cgccatccac aaggtgctgc
     3061 agctcatgct gcggcgcggc gagatccagc atcgcatgca gcgcaaggtt ctctaccgcc
     3121 tcaagagaat tctgcagtcg acggtaccgc gggcccggga tccaccggtc gccaccatgg
     3181 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     3241 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     3301 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     3361 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     3421 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     3481 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     3541 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     3601 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     3661 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     3721 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3781 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3841 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     3901 gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     3961 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     4021 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     4081 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     4141 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     4201 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4261 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4321 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4381 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4441 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4501 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4561 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4621 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4681 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4741 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4801 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4861 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4921 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4981 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     5041 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5101 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5161 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5221 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5281 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5341 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5401 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5461 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5521 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5581 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5641 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5701 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5761 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5821 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5881 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5941 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     6001 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     6061 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6121 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     6181 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6241 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6301 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6361 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6421 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6481 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6541 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6601 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6661 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6721 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6781 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6841 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6901 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6961 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7021 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7081 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     7141 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7201 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7261 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7321 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7381 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7441 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7501 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7561 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7621 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7681 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7741 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7801 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7861 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7921 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7981 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     8041 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8101 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8161 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8221 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8281 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8341 ctgacgt
//
