LOCUS       V020438                 8903 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020438
VERSION     V020438
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..4423
                     /label="THBS2(NM_003247)"
                     /note="THBS2(NM_003247)"
                     /gene="THBS2"
     polyA_signal    4470..4694
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4740..5168
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5182..5511
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5578..6372
                     /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    6546..6667
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6716..6732)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6740..6756
                     /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(6764..6794)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6809..6830
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7118..7703)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7874..8734)
                     /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(8735..8839)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gtctggaggc tggtcctgct ggctctgtgg gtgtggccca gcacgcaagc
      961 tggtcaccag gacaaagaca cgaccttcga ccttttcagt atcagcaaca tcaaccgcaa
     1021 gaccattggc gccaagcagt tccgcgggcc cgaccccggc gtgccggctt accgcttcgt
     1081 gcgctttgac tacatcccac cggtgaacgc agatgacctc agcaagatca ccaagatcat
     1141 gcggcagaag gagggcttct tcctcacggc ccagctcaag caggacggca agtccagggg
     1201 cacgctgttg gctctggagg gccccggtct ctcccagagg cagttcgaga tcgtctccaa
     1261 cggccccgcg gacacgctgg atctcaccta ctggattgac ggcacccggc atgtggtctc
     1321 cctggaggac gtcggcctgg ctgactcgca gtggaagaac gtcaccgtgc aggtggctgg
     1381 cgagacctac agcttgcacg tgggctgcga cctcatagac agcttcgctc tggacgagcc
     1441 cttctacgag cacctgcagg cggaaaagag ccggatgtac gtggccaaag gctctgccag
     1501 agagagtcac ttcaggggtt tgcttcagaa cgtccaccta gtgtttgaaa actctgtgga
     1561 agatattcta agcaagaagg gttgccagca aggccaggga gctgagatca acgccatcag
     1621 tgagaacaca gagacgctgc gcctgggtcc gcatgtcacc accgagtacg tgggccccag
     1681 ctcggagagg aggcccgagg tgtgcgaacg ctcgtgcgag gagctgggaa acatggtcca
     1741 ggagctctcg gggctccacg tcctcgtgaa ccagctcagc gagaacctca agagagtgtc
     1801 gaatgataac cagtttctct gggagctcat tggtggccct cctaagacaa ggaacatgtc
     1861 agcttgctgg caggatggcc ggttctttgc ggaaaatgaa acgtgggtgg tggacagctg
     1921 caccacgtgt acctgcaaga aatttaaaac catttgccac caaatcacct gcccgcctgc
     1981 aacctgcgcc agtccatcct ttgtggaagg cgaatgctgc ccttcctgcc tccactcggt
     2041 ggacggtgag gagggctggt ctccgtgggc agagtggacc cagtgctccg tgacgtgtgg
     2101 ctctgggacc cagcagagag gccggtcctg tgacgtcacc agcaacacct gcttggggcc
     2161 ctccatccag acacgggctt gcagtctgag caagtgtgac acccgcatcc ggcaggacgg
     2221 cggctggagc cactggtcac cttggtcttc atgctctgtg acctgtggag ttggcaatat
     2281 cacacgcatc cgtctctgca actccccagt gccccagatg gggggcaaga attgcaaagg
     2341 gagtggccgg gagaccaaag cctgccaggg cgccccatgc ccaatcgatg gccgctggag
     2401 cccctggtcc ccgtggtcgg cctgcactgt cacctgtgcc ggtgggatcc gggagcgcac
     2461 ccgggtctgc aacagccctg agcctcagta cggagggaag gcctgcgtgg gggatgtgca
     2521 ggagcgtcag atgtgcaaca agaggagctg ccccgtggat ggctgtttat ccaacccctg
     2581 cttcccggga gcccagtgca gcagcttccc cgatgggtcc tggtcatgcg gctcctgccc
     2641 tgtgggcttc ttgggcaatg gcacccactg tgaggacctg gacgagtgtg ccctggtccc
     2701 cgacatctgc ttctccacca gcaaggtgcc tcgctgtgtc aacactcagc ctggcttcca
     2761 ctgcctgccc tgcccgcccc gatacagagg gaaccagccc gtcggggtcg gcctggaagc
     2821 agccaagacg gaaaagcaag tgtgtgagcc cgaaaaccca tgcaaggaca agacacacaa
     2881 ctgccacaag cacgcggagt gcatctacct gggccacttc agcgacccca tgtacaagtg
     2941 cgagtgccag acaggctacg cgggcgacgg gctcatctgc ggggaggact cggacctgga
     3001 cggctggccc aacctcaatc tggtctgcgc caccaacgcc acctaccact gcatcaagga
     3061 taactgcccc catctgccaa attctgggca ggaagacttt gacaaggacg ggattggcga
     3121 tgcctgtgat gatgacgatg acaatgacgg tgtgaccgat gagaaggaca actgccagct
     3181 cctcttcaat ccccgccagg ctgactatga caaggatgag gttggggacc gctgtgacaa
     3241 ctgcccttac gtgcacaacc ctgcccagat cgacacagac aacaatggag agggtgacgc
     3301 ctgctccgtg gacattgatg gggacgatgt cttcaatgaa cgagacaatt gtccctacgt
     3361 ctacaacact gaccagaggg acacggatgg tgacggtgtg ggggatcact gtgacaactg
     3421 ccccctggtg cacaaccctg accagaccga cgtggacaat gaccttgttg gggaccagtg
     3481 tgacaacaac gaggacatag atgacgacgg ccaccagaac aaccaggaca actgccccta
     3541 catctccaac gccaaccagg ctgaccatga cagagacggc cagggcgacg cctgtgaccc
     3601 tgatgatgac aacgatggcg tccccgatga cagggacaac tgccggcttg tgttcaaccc
     3661 agaccaggag gacttggacg gtgatggacg gggtgatatt tgtaaagatg attttgacaa
     3721 tgacaacatc ccagatattg atgatgtgtg tcctgaaaac aatgccatca gtgagacaga
     3781 cttcaggaac ttccagatgg tccccttgga tcccaaaggg accacccaaa ttgatcccaa
     3841 ctgggtcatt cgccatcaag gcaaggagct ggttcagaca gccaactcgg accccggcat
     3901 cgctgtaggt tttgacgagt ttgggtctgt ggacttcagt ggcacattct acgtaaacac
     3961 tgaccgggac gacgactatg ccggcttcgt ctttggttac cagtcaagca gccgcttcta
     4021 tgtggtgatg tggaagcagg tgacgcagac ctactgggag gaccagccca cgcgggccta
     4081 tggctactcc ggcgtgtccc tcaaggtggt gaactccacc acggggacgg gcgagcacct
     4141 gaggaacgcg ctgtggcaca cggggaacac gccggggcag gtgcgaacct tatggcacga
     4201 ccccaggaac attggctgga aggactacac ggcctatagg tggcacctga ctcacaggcc
     4261 caagactggc tacatcagag tcttagtgca tgaaggaaaa caggtcatgg cagactcagg
     4321 acctatctat gaccaaacct acgctggcgg gcggctgggt ctatttgtct tctctcaaga
     4381 aatggtctat ttctcagacc tcaagtacga atgcagagat atttaactcg agtctagagg
     4441 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     4501 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     4561 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     4621 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     4681 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     4741 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4801 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4861 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4921 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4981 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5041 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5101 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5161 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     5221 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     5281 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     5341 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5401 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     5461 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     5521 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     5581 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5641 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     5701 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     5761 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5821 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5881 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5941 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     6001 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     6061 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     6121 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     6181 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     6241 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     6301 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     6361 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     6421 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     6481 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     6541 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6601 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6661 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     6721 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6781 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6841 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6901 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6961 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     7021 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     7081 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7141 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7201 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7261 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7321 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7381 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7441 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7501 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     7561 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7621 cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta
     7681 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7741 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7801 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7861 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7921 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7981 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8041 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8101 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8161 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8221 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8281 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8341 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8401 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8461 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8521 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8581 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8641 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8701 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8761 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8821 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     8881 gagatctccc gatcccctat ggt
//
