LOCUS       V018695                 8246 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018695
VERSION     V018695
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             932..3721
                     /label="ADAMTS5(NM_007038)"
                     /note="ADAMTS5(NM_007038)"
                     /gene="ADAMTS5"
     CDS             3743..3808
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3846..4070
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4116..4544
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4558..4887
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4954..5748
                     /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    5922..6043
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6092..6108)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6116..6132
                     /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(6140..6170)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6185..6206
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6494..7079)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7250..8110)
                     /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(8111..8215)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccgccac catgctgctc gggtgggcgt ccctgctgct
      961 gtgcgcgttc cgcctgcccc tggccgcggt cggccccgcc gcgacacctg cccaggataa
     1021 agccgggcag cctccgactg ctgcagcagc cgcccagccc cgccggcggc agggggagga
     1081 ggtgcaggag cgagccgagc ctcccggcca cccgcacccc ctggcgcagc ggcgcaggag
     1141 caaggggctg gtgcagaaca tcgaccaact ctactccggc ggcggcaagg tgggctacct
     1201 cgtctacgcg ggcggccgga ggttcctctt ggacctggag cgagatggtt cggtgggcat
     1261 tgctggcttc gtgcccgcag gaggcgggac gagtgcgccc tggcgccacc ggagccactg
     1321 cttctatcgg ggcacagtgg acggtagtcc ccgctctctg gctgtctttg acctctgtgg
     1381 gggtctcgac ggcttcttcg cggtcaagca cgcgcgctac accctaaagc cactgctgcg
     1441 cggaccctgg gcggaggaag aaaaggggcg cgtgtacggg gatgggtccg cacggatcct
     1501 gcacgtctac acccgcgagg gcttcagctt cgaggccctg ccgccgcgcg ccagctgcga
     1561 aacccccgcg tccacaccgg aggcccacga gcatgctccg gcgcacagca acccgagcgg
     1621 acgcgcagca ctggcctcgc agctcttgga ccagtccgct ctctcgcccg ctgggggctc
     1681 aggaccgcag acgtggtggc ggcggcggcg ccgctccatc tcccgggccc gccaggtgga
     1741 gctgcttctg gtggctgacg cgtccatggc gcggttgtat ggccggggcc tgcagcatta
     1801 cctgctgacc ctggcctcca tcgccaatag gctgtacagc catgctagca tcgagaacca
     1861 catccgcctg gccgtggtga aggtggtggt gctaggcgac aaggacaaga gcctggaagt
     1921 gagcaagaac gctgccacca cactcaagaa cttttgcaag tggcagcacc aacacaacca
     1981 gctgggagat gaccatgagg agcactacga tgcagctatc ctgtttactc gggaggattt
     2041 atgtgggcat cattcatgtg acaccctggg aatggcagac gttgggacca tatgttctcc
     2101 agagcgcagc tgtgctgtga ttgaagacga tggcctccac gcagccttca ctgtggctca
     2161 cgaaatcgga catttacttg gcctctccca tgacgattcc aaattctgtg aagagacctt
     2221 tggttccaca gaagataagc gcttaatgtc ttccatcctt accagcattg atgcatctaa
     2281 gccctggtcc aaatgcactt cagccaccat cacagaattc ctggatgatg gccatggtaa
     2341 ctgtttgctg gacctaccac gaaagcagat cctgggcccc gaagaactcc caggacagac
     2401 ctacgatgcc acccagcagt gcaacctgac attcgggcct gagtactccg tgtgtcccgg
     2461 catggatgtc tgtgctcgcc tgtggtgtgc tgtggtacgc cagggccaga tggtctgtct
     2521 gaccaagaag ctgcctgcgg tggaagggac gccttgtgga aaggggagaa tctgcctgca
     2581 gggcaaatgt gtggacaaaa ccaagaaaaa atattattca acgtcaagcc atggcaactg
     2641 gggatcttgg ggatcctggg gccagtgttc tcgctcatgt ggaggaggag tgcagtttgc
     2701 ctatcgtcac tgtaataacc ctgctcccag aaacaacgga cgctactgca cagggaagag
     2761 ggccatctac cgctcctgca gtctcatgcc ctgcccaccc aatggtaaat catttcgtca
     2821 tgaacagtgt gaggccaaaa atggctatca gtctgatgca aaaggagtca aaacttttgt
     2881 ggaatgggtt cccaaatatg caggtgtcct gccagcggat gtgtgcaagc tgacctgcag
     2941 agccaagggc actggctact atgtggtatt ttctccaaag gtgaccgatg gcactgaatg
     3001 taggctgtac agtaattccg tctgcgtccg ggggaagtgt gtgagaactg gctgtgacgg
     3061 catcattggc tcaaagctgc agtatgacaa gtgcggagta tgtggaggag acaactccag
     3121 ctgtacaaag attgttggaa cctttaataa gaaaagtaag ggttacactg acgtggtgag
     3181 gattcctgaa ggggcaaccc acataaaagt tcgacagttc aaagccaaag accagactag
     3241 attcactgcc tatttagccc tgaaaaagaa aaacggtgag taccttatca atggaaagta
     3301 catgatctcc acttcagaga ctatcattga catcaatgga acagtcatga actatagcgg
     3361 ttggagccac agggatgact tcctgcatgg catgggctac tctgccacga aggaaattct
     3421 aatagtgcag attcttgcaa cagaccccac taaaccatta gatgtccgtt atagcttttt
     3481 tgttcccaag aagtccactc caaaagtaaa ctctgtcact agtcatggca gcaataaagt
     3541 gggatcacac acttcgcagc cgcagtgggt cacgggccca tggctcgcct gctctaggac
     3601 ctgtgacaca ggttggcaca ccagaacggt gcagtgccag gatggaaacc ggaagttagc
     3661 aaaaggatgt cctctctccc aaaggccttc tgcgtttaag caatgcttgt tgaagaaatg
     3721 tctcgagtct agagggccct tcgactacaa agaccatgac ggtgattata aagatcatga
     3781 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc
     3841 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3901 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3961 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     4021 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     4081 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     4141 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4201 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4261 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4321 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4381 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4441 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4501 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     4561 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4621 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4681 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4741 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4801 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4861 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     4921 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4981 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5041 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5101 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5161 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5221 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5281 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5341 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5401 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5461 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5521 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5581 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5641 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5701 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5761 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5821 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5881 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     5941 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6001 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     6061 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6121 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6181 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6241 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6301 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6361 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6421 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6481 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6541 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6601 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6661 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6721 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6781 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6841 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6901 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6961 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7021 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7081 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7141 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7201 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7261 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7321 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7381 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7441 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7501 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7561 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7621 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7681 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7741 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7801 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7861 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7921 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7981 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8041 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8101 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8161 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8221 ttccccgaaa agtgccacct gacgtc
//
