LOCUS       V024573                 9426 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024573
VERSION     V024573
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..4106
                     /label="ADAMTS6(NM_197941)"
                     /note="ADAMTS6(NM_197941)"
                     /gene="ADAMTS6"
     CDS             4113..4178
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4202..4790
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4856..4904
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5106..5534
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5548..5877
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5944..6738
                     /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    6914..7035
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7084..7100)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7108..7124
                     /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(7132..7162)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7177..7198
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7486..8074)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8245..9105)
                     /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(9106..9210)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgga aattttgtgg aagacgttga
      781 cctggatttt gagcctcatc atggcttcat cggaatttca tagtgaccac aggctttcat
      841 acagttctca agaggaattc ctgacttatc ttgaacacta ccagctaact attccaataa
      901 gggttgatca aaatggagca tttctcagct ttactgtgaa aaatgataaa cactcaagga
      961 gaagacggag tatggaccct attgatccac agcaggcagt atctaagtta ttttttaaac
     1021 tttcagccta tggcaagcac tttcatctaa acttgactct caacacagat tttgtgtcca
     1081 aacattttac agtagaatat tgggggaaag atggacccca gtggaaacat gattttttag
     1141 acaactgtca ttacacagga tatttgcaag atcaacgtag tacaactaaa gtggctttaa
     1201 gcaactgtgt tgggttgcat ggtgttattg ctacagaaga tgaagagtat tttatcgaac
     1261 ctttaaagaa taccacagag gattccaagc attttagtta tgaaaatggc caccctcatg
     1321 ttatttacaa aaagtctgcc cttcaacaac gacatctgta tgatcactct cattgtgggg
     1381 tttcggattt cacaagaagt ggcaaacctt ggtggctgaa tgacacatcc actgtttctt
     1441 attcactacc aattaacaac acacatatcc accacagaca gaagagatca gtgagcattg
     1501 aacggtttgt ggagacattg gtagtggcag acaaaatgat ggtgggctac catggccgca
     1561 aagacattga acattacatt ttgagtgtga tgaatattgt tgccaaactt taccgtgatt
     1621 ccagcctagg aaacgttgtg aatattatag tggcccgctt aattgttctc acagaagatc
     1681 agccaaactt ggagataaac caccatgcag acaagtccct cgatagcttc tgtaaatggc
     1741 agaaatccat tctctcccac caaagtgatg gaaacaccat tccagaaaat gggattgccc
     1801 accacgataa tgcagttctt attactagat atgatatctg cacttataaa aataagccct
     1861 gtggaacact gggcttggcc tctgtggctg gaatgtgtga gcctgaaagg agctgcagca
     1921 ttaatgaaga cattggcctg ggttcagctt ttaccattgc acatgagatt ggtcacaatt
     1981 ttggtatgaa ccatgatgga attggaaatt cttgtgggac gaaaggtcat gaagcagcaa
     2041 aacttatggc agctcacatt actgcgaata ccaatccttt ttcctggtct gcttgcagtc
     2101 gagactacat caccagcttt ctagattcag gccgtggtac ttgccttgat aatgagcctc
     2161 ccaagcgtga ctttctttat ccagctgtgg ccccaggtca ggtgtatgat gctgatgagc
     2221 aatgtcgttt ccagtatgga gcaacctccc gccaatgtaa atatggggaa gtgtgtagag
     2281 agctctggtg tctcagcaaa agcaaccgct gtgtcaccaa cagtattcca gcagctgagg
     2341 ggacactgtg tcaaactggg aatattgaaa aagggtggtg ttatcaggga gattgtgttc
     2401 cttttggcac ttggccccag agcatagatg ggggctgggg tccctggtca ctatggggag
     2461 agtgcagcag gacctgcggg ggaggcgtct cctcatccct aagacactgt gacagtccag
     2521 caccttcagg aggtggaaaa tattgccttg gggaaaggaa acggtatcgc tcctgtaaca
     2581 cagatccatg ccctttgggt tcccgagatt ttcgagagaa acagtgtgca gactttgaca
     2641 atatgccttt ccgaggaaag tattataact ggaaacccta tactggaggt ggggtaaaac
     2701 cttgtgcatt aaactgcttg gctgaaggtt ataatttcta cactgaacgt gctcctgcgg
     2761 tgatcgatgg gacccagtgc aatgcggatt cactggatat ctgcatcaat ggagaatgca
     2821 agcacgtagg ctgtgataat attttgggat ctgatgctag ggaagataga tgtcgagtct
     2881 gtggagggga cggaagcaca tgtgatgcca ttgaagggtt cttcaatgat tcactgccca
     2941 ggggaggcta catggaagtg gtgcagatac caagaggctc tgttcacatt gaagttagag
     3001 aagttgccat gtcaaagaac tatattgctt taaaatctga aggagatgat tactatatta
     3061 atggtgcctg gactattgac tggcctagga aatttgatgt tgctgggaca gcttttcatt
     3121 acaagagacc aactgatgaa ccagaatcct tggaagctct aggtcctacc tcagaaaatc
     3181 tcatcgtcat ggttctgctt caagaacaga atttgggaat taggtataag ttcaatgttc
     3241 ccatcactcg aactggcagt ggagataatg aagttggctt tacatggaat catcagcctt
     3301 ggtcagaatg ctcagctact tgtgctggag gtgtccaaag acaggaggtg gtctgtaaaa
     3361 ggttggatga caactccatt gtccagaaca attactgtga tcctgacagt aagccacctg
     3421 aaaatcaaag agcctgcaac actgagccct gcccacctga gtggttcatt ggggattggt
     3481 tggaatgcag caagacttgt gatggtggga tgcgcacaag ggcagtgctc tgcatcagga
     3541 agatcggacc ttctgaggag gagacgctgg actacagtgg ttgtttaaca caccggcctg
     3601 tcgaaaaaga gccctgcaac aaccagtcat gtccaccaca gtgggtggct ttggactggt
     3661 ctgagtgtac tccaaaatgt ggtccaggat tcaagcatcg gattgttctg tgcaagagca
     3721 gtgacctttc taagacattc ccagctgcac aatgtccaga ggaaagcaaa cctcctgtcc
     3781 gcatccgctg cagtttgggc cgctgccctc ctcctcgctg ggtcacagga gactggggcc
     3841 agtgttctgc tcagtgtggc cttggacagc agatgagaac tgtgcagtgt ctctcctaca
     3901 ccggacaggc atctagtgac tgtctagaaa ctgttcggcc tccatcaatg cagcagtgtg
     3961 aaagcaaatg tgacagtacc cccatttcta atactgaaga gtgcaaagat gtgaataaag
     4021 tggcttattg cccactggtg ctgaagttca agttctgcag tcgagcatac ttcagacaga
     4081 tgtgttgtaa gacctgccaa ggacacggat ccgactacaa agaccatgac ggtgattata
     4141 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     4201 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     4261 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     4321 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     4381 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     4441 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     4501 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     4561 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     4621 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     4681 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     4741 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     4801 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     4861 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     4921 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4981 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     5041 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     5101 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5161 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5221 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     5281 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5341 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5401 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5461 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5521 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     5581 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     5641 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5701 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5761 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     5821 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     5881 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     5941 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     6001 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     6061 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     6121 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     6181 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     6241 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     6301 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     6361 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     6421 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     6481 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6541 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6601 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6661 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6721 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     6781 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     6841 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6901 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6961 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     7021 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     7081 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     7141 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     7201 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     7261 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     7321 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     7381 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7441 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7501 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7561 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7621 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7681 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7741 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7801 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7861 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7921 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7981 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     8041 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     8101 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     8161 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     8221 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     8281 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     8341 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     8401 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     8461 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     8521 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     8581 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     8641 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8701 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8761 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8821 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8881 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8941 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     9001 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     9061 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     9121 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     9181 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     9241 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     9301 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     9361 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     9421 cttagg
//
