LOCUS       V018538                 9500 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018538
VERSION     V018538
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"
     CDS             671..4180
                     /label="THBS1(NM_003246)"
                     /note="THBS1(NM_003246)"
                     /gene="THBS1"
     CDS             4187..4252
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4276..4864
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4930..4978
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5180..5608
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5622..5951
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6018..6812
                     /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    6988..7109
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7158..7174)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7182..7198
                     /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(7206..7236)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7251..7272
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7560..8148)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8319..9179)
                     /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(9180..9284)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgccacc atggggctgg cctggggact aggcgtcctg ttcctgatgc atgtgtgtgg
      721 caccaaccgc attccagagt ctggcggaga caacagcgtg tttgacatct ttgaactcac
      781 cggggccgcc cgcaaggggt ctgggcgccg actggtgaag ggccccgacc cttccagccc
      841 agctttccgc atcgaggatg ccaacctgat cccccctgtg cctgatgaca agttccaaga
      901 cctggtggat gctgtgcggg cagaaaaggg tttcctcctt ctggcatccc tgaggcagat
      961 gaagaagacc cggggcacgc tgctggccct ggagcggaaa gaccactctg gccaggtctt
     1021 cagcgtggtg tccaatggca aggcgggcac cctggacctc agcctgaccg tccaaggaaa
     1081 gcagcacgtg gtgtctgtgg aagaagctct cctggcaacc ggccagtgga agagcatcac
     1141 cctgtttgtg caggaagaca gggcccagct gtacatcgac tgtgaaaaga tggagaatgc
     1201 tgagttggac gtccccatcc aaagcgtctt caccagagac ctggccagca tcgccagact
     1261 ccgcatcgca aaggggggcg tcaatgacaa tttccagggg gtgctgcaga atgtgaggtt
     1321 tgtctttgga accacaccag aagacatcct caggaacaaa ggctgctcca gctctaccag
     1381 tgtcctcctc acccttgaca acaacgtggt gaatggttcc agccctgcca tccgcactaa
     1441 ctacattggc cacaagacaa aggacttgca agccatctgc ggcatctcct gtgatgagct
     1501 gtccagcatg gtcctggaac tcaggggcct gcgcaccatt gtgaccacgc tgcaggacag
     1561 catccgcaaa gtgactgaag agaacaaaga gttggccaat gagctgaggc ggcctcccct
     1621 atgctatcac aacggagttc agtacagaaa taacgaggaa tggactgttg atagctgcac
     1681 tgagtgtcac tgtcagaact cagttaccat ctgcaaaaag gtgtcctgcc ccatcatgcc
     1741 ctgctccaat gccacagttc ctgatggaga atgctgtcct cgctgttggc ccagcgactc
     1801 tgcggacgat ggctggtctc catggtccga gtggacctcc tgttctacga gctgtggcaa
     1861 tggaattcag cagcgcggcc gctcctgcga tagcctcaac aaccgatgtg agggctcctc
     1921 ggtccagaca cggacctgcc acattcagga gtgtgacaag agatttaaac aggatggtgg
     1981 ctggagccac tggtccccgt ggtcatcttg ttctgtgaca tgtggtgatg gtgtgatcac
     2041 aaggatccgg ctctgcaact ctcccagccc ccagatgaac gggaaaccct gtgaaggcga
     2101 agcgcgggag accaaagcct gcaagaaaga cgcctgcccc atcaatggag gctggggtcc
     2161 ttggtcacca tgggacatct gttctgtcac ctgtggagga ggggtacaga aacgtagtcg
     2221 tctctgcaac aaccccacac cccagtttgg aggcaaggac tgcgttggtg atgtaacaga
     2281 aaaccagatc tgcaacaagc aggactgtcc aattgatgga tgcctgtcca atccctgctt
     2341 tgccggcgtg aagtgtacta gctaccctga tggcagctgg aaatgtggtg cttgtccccc
     2401 tggttacagt ggaaatggca tccagtgcac agatgttgat gagtgcaaag aagtgcctga
     2461 tgcctgcttc aaccacaatg gagagcaccg gtgtgagaac acggaccccg gctacaactg
     2521 cctgccctgc cccccacgct tcaccggctc acagcccttc ggccagggtg tcgaacatgc
     2581 cacggccaac aaacaggtgt gcaagccccg taacccctgc acggatggga cccacgactg
     2641 caacaagaac gccaagtgca actacctggg ccactatagc gaccccatgt accgctgcga
     2701 gtgcaagcct ggctacgctg gcaatggcat catctgcggg gaggacacag acctggatgg
     2761 ctggcccaat gagaacctgg tgtgcgtggc caatgcgact taccactgca aaaaggataa
     2821 ttgccccaac cttcccaact cagggcagga agactatgac aaggatggaa ttggtgatgc
     2881 ctgtgatgat gacgatgaca atgataaaat tccagatgac agggacaact gtccattcca
     2941 ttacaaccca gctcagtatg actatgacag agatgatgtg ggagaccgct gtgacaactg
     3001 tccctacaac cacaacccag atcaggcaga cacagacaac aatggggaag gagacgcctg
     3061 tgctgcagac attgatggag acggtatcct caatgaacgg gacaactgcc agtacgtcta
     3121 caatgtggac cagagagaca ctgatatgga tggggttgga gatcagtgtg acaattgccc
     3181 cttggaacac aatccggatc agctggactc tgactcagac cgcattggag atacctgtga
     3241 caacaatcag gatattgatg aagatggcca ccagaacaat ctggacaact gtccctatgt
     3301 gcccaatgcc aaccaggctg accatgacaa agatggcaag ggagatgcct gtgaccacga
     3361 tgatgacaac gatggcattc ctgatgacaa ggacaactgc agactcgtgc ccaatcccga
     3421 ccagaaggac tctgacggcg atggtcgagg tgatgcctgc aaagatgatt ttgaccatga
     3481 cagtgtgcca gacatcgatg acatctgtcc tgagaatgtt gacatcagtg agaccgattt
     3541 ccgccgattc cagatgattc ctctggaccc caaagggaca tcccaaaatg accctaactg
     3601 ggttgtacgc catcagggta aagaactcgt ccagactgtc aactgtgatc ctggactcgc
     3661 tgtaggttat gatgagttta atgctgtgga cttcagtggc accttcttca tcaacaccga
     3721 aagggacgat gactatgctg gatttgtctt tggctaccag tccagcagcc gcttttatgt
     3781 tgtgatgtgg aagcaagtca cccagtccta ctgggacacc aaccccacga gggctcaggg
     3841 atactcgggc ctttctgtga aagttgtaaa ctccaccaca gggcctggcg agcacctgcg
     3901 gaacgccctg tggcacacag gaaacacccc tggccaggtg cgcaccctgt ggcatgaccc
     3961 tcgtcacata ggctggaaag atttcaccgc ctacagatgg cgtctcagcc acaggccaaa
     4021 gacgggtttc attagagtgg tgatgtatga agggaagaaa atcatggctg actcaggacc
     4081 catctatgat aaaacctatg ctggtggtag actagggttg tttgtcttct ctcaagaaat
     4141 ggtgttcttc tctgacctga aatacgaatg tagagatccc ctcgaggact acaaagacca
     4201 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg
     4261 gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     4321 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     4381 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     4441 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     4501 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     4561 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     4621 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     4681 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     4741 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     4801 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     4861 ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4921 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat
     4981 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5041 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5101 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg
     5161 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5221 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5281 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc
     5341 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5401 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5461 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5521 gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag
     5581 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     5641 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5701 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5761 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5821 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5881 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5941 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     6001 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     6061 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     6121 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     6181 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     6241 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     6301 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     6361 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     6421 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     6481 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     6541 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     6601 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     6661 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     6721 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     6781 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc
     6841 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     6901 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     6961 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     7021 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     7081 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     7141 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     7201 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     7261 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     7321 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     7381 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     7441 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     7501 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7561 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7621 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7681 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7741 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7801 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7861 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7921 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7981 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     8041 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     8101 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     8161 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     8221 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     8281 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     8341 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     8401 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     8461 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8521 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8581 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     8641 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8701 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8761 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8821 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8881 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8941 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     9001 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     9061 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     9121 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     9181 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     9241 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     9301 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta
     9361 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     9421 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     9481 gcatgaagaa tctgcttagg
//
