LOCUS       V022057                 9731 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022057
VERSION     V022057
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             941..5221
                     /label="ADAMTS13(NM_139025)"
                     /note="ADAMTS13(NM_139025)"
                     /gene="ADAMTS13"
     CDS             5228..5293
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5331..5555
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5601..6029
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6043..6372
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6439..7233
                     /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    7407..7528
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7577..7593)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7601..7617
                     /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(7625..7655)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7670..7691
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7979..8564)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8735..9595)
                     /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(9596..9700)
                     /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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgcaccagc gtcacccccg
      961 ggcaagatgc cctcccctct gtgtggccgg aatccttgcc tgtggctttc tcctgggctg
     1021 ctggggaccc tcccatttcc agcagagttg tcttcaggct ttggagccac aggccgtgtc
     1081 ttcttacttg agccctggtg ctcccttaaa aggccgccct ccttcccctg gcttccagag
     1141 gcagaggcag aggcagaggc gggctgcagg cggcatccta cacctggagc tgctggtggc
     1201 cgtgggcccc gatgtcttcc aggctcacca ggaggacaca gagcgctatg tgctcaccaa
     1261 cctcaacatc ggggcagaac tgcttcggga cccgtccctg ggggctcagt ttcgggtgca
     1321 cctggtgaag atggtcattc tgacagagcc tgagggtgcc ccaaatatca cagccaacct
     1381 cacctcgtcc ctgctgagcg tctgtgggtg gagccagacc atcaaccctg aggacgacac
     1441 ggatcctggc catgctgacc tggtcctcta tatcactagg tttgacctgg agttgcctga
     1501 tggtaaccgg caggtgcggg gcgtcaccca gctgggcggt gcctgctccc caacctggag
     1561 ctgcctcatt accgaggaca ctggcttcga cctgggagtc accattgccc atgagattgg
     1621 gcacagcttc ggcctggagc acgacggcgc gcccggcagc ggctgcggcc ccagcggaca
     1681 cgtgatggct tcggacggcg ccgcgccccg cgccggcctc gcctggtccc cctgcagccg
     1741 ccggcagctg ctgagcctgc tcagcgcagg acgggcgcgc tgcgtgtggg acccgccgcg
     1801 gcctcaaccc gggtccgcgg ggcacccgcc ggatgcgcag cctggcctct actacagcgc
     1861 caacgagcag tgccgcgtgg ccttcggccc caaggctgtc gcctgcacct tcgccaggga
     1921 gcacctggat atgtgccagg ccctctcctg ccacacagac ccgctggacc aaagcagctg
     1981 cagccgcctc ctcgttcctc tcctggatgg gacagaatgt ggcgtggaga agtggtgctc
     2041 caagggtcgc tgccgctccc tggtggagct gacccccata gcagcagtgc atgggcgctg
     2101 gtctagctgg ggtccccgaa gtccttgctc ccgctcctgc ggaggaggtg tggtcaccag
     2161 gaggcggcag tgcaacaacc ccagacctgc ctttgggggg cgtgcatgtg ttggtgctga
     2221 cctccaggcc gagatgtgca acactcaggc ctgcgagaag acccagctgg agttcatgtc
     2281 gcaacagtgc gccaggaccg acggccagcc gctgcgctcc tcccctggcg gcgcctcctt
     2341 ctaccactgg ggtgctgctg taccacacag ccaaggggat gctctgtgca gacacatgtg
     2401 ccgggccatt ggcgagagct tcatcatgaa gcgtggagac agcttcctcg atgggacccg
     2461 gtgtatgcca agtggccccc gggaggacgg gaccctgagc ctgtgtgtgt cgggcagctg
     2521 caggacattt ggctgtgatg gtaggatgga ctcccagcag gtatgggaca ggtgccaggt
     2581 gtgtggtggg gacaacagca cgtgcagccc acggaagggc tctttcacag ctggcagagc
     2641 gagagaatat gtcacatttc tgacagttac ccccaacctg accagtgtct acattgccaa
     2701 ccacaggcct ctcttcacac acttggcggt gaggatcgga gggcgctatg tcgtggctgg
     2761 gaagatgagc atctccccta acaccaccta cccctccctc ctggaggatg gtcgtgtcga
     2821 gtacagagtg gccctcaccg aggaccggct gccccgcctg gaggagatcc gcatctgggg
     2881 acccctccag gaagatgctg acatccaggt ttacaggcgg tatggcgagg agtatggcaa
     2941 cctcacccgc ccagacatca ccttcaccta cttccagcct aagccacggc aggcctgggt
     3001 gtgggccgct gtgcgtgggc cctgctcggt gagctgtggg gcagggctgc gctgggtaaa
     3061 ctacagctgc ctggaccagg ccaggaagga gttggtggag actgtccagt gccaagggag
     3121 ccagcagcca ccagcgtggc cagaggcctg cgtgctcgaa ccctgccctc cctactgggc
     3181 ggtgggagac ttcggcccat gcagcgcctc ctgtgggggc ggcctgcggg agcggccagt
     3241 gcgctgcgtg gaggcccagg gcagcctcct gaagacattg cccccagccc ggtgcagagc
     3301 aggggcccag cagccagctg tggcgctgga aacctgcaac ccccagccct gccctgccag
     3361 gtgggaggtg tcagagccca gctcatgcac atcagctggt ggagcaggcc tggccttgga
     3421 gaacgagacc tgtgtgccag gggcagatgg cctggaggct ccagtgactg aggggcctgg
     3481 ctccgtagat gagaagctgc ctgcccctga gccctgtgtc gggatgtcat gtcctccagg
     3541 ctggggccat ctggatgcca cctctgcagg ggagaaggct ccctccccat ggggcagcat
     3601 caggacgggg gctcaagctg cacacgtgtg gacccctgcg gcagggtcgt gctccgtctc
     3661 ctgcgggcga ggtctgatgg agctgcgttt cctgtgcatg gactctgccc tcagggtgcc
     3721 tgtccaggaa gagctgtgtg gcctggcaag caagcctggg agccggcggg aggtctgcca
     3781 ggctgtcccg tgccctgctc ggtggcagta caagctggcg gcctgcagcg tgagctgtgg
     3841 gagaggggtc gtgcggagga tcctgtattg tgcccgggcc catggggagg acgatggtga
     3901 ggagatcctg ttggacaccc agtgccaggg gctgcctcgc ccggaacccc aggaggcctg
     3961 cagcctggag ccctgcccac ctaggtggaa agtcatgtcc cttggcccat gttcggccag
     4021 ctgtggcctt ggcactgcta gacgctcggt ggcctgtgtg cagctcgacc aaggccagga
     4081 cgtggaggtg gacgaggcgg cctgtgcggc gctggtgcgg cccgaggcca gtgtcccctg
     4141 tctcattgcc gactgcacct accgctggca tgttggcacc tggatggagt gctctgtttc
     4201 ctgtggggat ggcatccagc gccggcgtga cacctgcctc ggaccccagg cccaggcgcc
     4261 tgtgccagct gatttctgcc agcacttgcc caagccggtg actgtgcgtg gctgctgggc
     4321 tgggccctgt gtgggacagg gtacgcccag cctggtgccc cacgaagaag ccgctgctcc
     4381 aggacggacc acagccaccc ctgctggtgc ctccctggag tggtcccagg cccggggcct
     4441 gctcttctcc ccggctcccc agcctcggcg gctcctgccc gggccccagg aaaactcagt
     4501 gcagtccagt gcctgtggca ggcagcacct tgagccaaca ggaaccattg acatgcgagg
     4561 cccagggcag gcagactgtg cagtggccat tgggcggccc ctcggggagg tggtgaccct
     4621 ccgcgtcctt gagagttctc tcaactgcag tgcgggggac atgttgctgc tttggggccg
     4681 gctcacctgg aggaagatgt gcaggaagct gttggacatg actttcagct ccaagaccaa
     4741 cacgctggtg gtgaggcagc gctgcgggcg gccaggaggt ggggtgctgc tgcggtatgg
     4801 gagccagctt gctcctgaaa ccttctacag agaatgtgac atgcagctct ttgggccctg
     4861 gggtgaaatc gtgagcccct cgctgagtcc agccacgagt aatgcagggg gctgccggct
     4921 cttcattaat gtggctccgc acgcacggat tgccatccat gccctggcca ccaacatggg
     4981 cgctgggacc gagggagcca atgccagcta catcttgatc cgggacaccc acagcttgag
     5041 gaccacagcg ttccatgggc agcaggtgct ctactgggag tcagagagca gccaggctga
     5101 gatggagttc agcgagggct tcctgaaggc tcaggccagc ctgcggggcc agtactggac
     5161 cctccaatca tgggtaccgg agatgcagga ccctcagtcc tggaagggaa aggaaggaac
     5221 cctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     5281 tgacgatgac aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     5341 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     5401 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     5461 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     5521 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     5581 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5641 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5701 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5761 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5821 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5881 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5941 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     6001 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     6061 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     6121 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     6181 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     6241 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6301 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     6361 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     6421 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     6481 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     6541 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     6601 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     6661 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6721 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6781 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6841 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6901 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6961 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     7021 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     7081 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     7141 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     7201 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     7261 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     7321 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     7381 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     7441 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     7501 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     7561 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     7621 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     7681 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     7741 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     7801 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     7861 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     7921 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7981 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     8041 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     8101 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     8161 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     8221 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     8281 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     8341 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     8401 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     8461 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt
     8521 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     8581 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     8641 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     8701 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     8761 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8821 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8881 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8941 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     9001 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     9061 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     9121 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     9181 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     9241 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     9301 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     9361 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     9421 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     9481 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     9541 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     9601 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     9661 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     9721 cacctgacgt c
//
