LOCUS       V018394                 7349 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018394
VERSION     V018394
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        61..364
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        365..568
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             625..3135
                     /label="ADAMTS4(NM_005099)"
                     /note="ADAMTS4(NM_005099)"
                     /gene="ADAMTS4"
     CDS             3160..3177
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /transl_table=1
                     /translation="LVPRGS"
                     /label="thrombin site"
     CDS             3190..3273
                     /codon_start=1
                     /product="two Strep-Tag®II moieties connected by a linker
                     for enhanced binding to Strep-Tactin®,&nbsp;an engineered
                     form of streptavidin (Schmidt et al., 2013)"
                     /transl_table=1
                     /translation="WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK"
                     /label="Twin-Strep-tag"
     CDS             3295..4014
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     polyA_signal    4137..4258
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(4265..4720)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4747..4851
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        4853..5210
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5245..6039
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6271..6318
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6647..7235
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgaggc caccatgtcc cagacaggct cgcatcccgg gaggggcttg
      661 gcagggcgct ggctgtgggg agcccaaccc tgcctcctgc tccccattgt gccgctctcc
      721 tggctggtgt ggctgcttct gctactgctg gcctctctcc tgccctcagc ccggctggcc
      781 agccccctcc cccgggagga ggagatcgtg tttccagaga agctcaacgg cagcgtcctg
      841 cctggctcgg gcgcccctgc caggctgttg tgccgcttgc aggcctttgg ggagacgctg
      901 ctactagagc tggagcagga ctccggtgtg caggtcgagg ggctgacagt gcagtacctg
      961 ggccaggcgc ctgagctgct gggtggagca gagcctggca cctacctgac tggcaccatc
     1021 aatggagatc cggagtcggt ggcatctctg cactgggatg ggggagccct gttaggcgtg
     1081 ttacaatatc ggggggctga actccacctc cagcccctgg agggaggcac ccctaactct
     1141 gctgggggac ctggggctca catcctacgc cggaagagtc ctgccagcgg tcaaggtccc
     1201 atgtgcaacg tcaaggctcc tcttggaagc cccagcccca gaccccgaag agccaagcgc
     1261 tttgcttcac tgagtagatt tgtggagaca ctggtggtgg cagatgacaa gatggccgca
     1321 ttccacggtg cggggctaaa gcgctacctg ctaacagtga tggcagcagc agccaaggcc
     1381 ttcaagcacc caagcatccg caatcctgtc agcttggtgg tgactcggct agtgatcctg
     1441 gggtcaggcg aggaggggcc ccaagtgggg cccagtgctg cccagaccct gcgcagcttc
     1501 tgtgcctggc agcggggcct caacacccct gaggactcgg accctgacca ctttgacaca
     1561 gccattctgt ttacccgtca ggacctgtgt ggagtctcca cttgcgacac gctgggtatg
     1621 gctgatgtgg gcaccgtctg tgacccggct cggagctgtg ccattgtgga ggatgatggg
     1681 ctccagtcag ccttcactgc tgctcatgaa ctgggtcatg tcttcaacat gctccatgac
     1741 aactccaagc catgcatcag tttgaatggg cctttgagca cctctcgcca tgtcatggcc
     1801 cctgtgatgg ctcatgtgga tcctgaggag ccctggtccc cctgcagtgc ccgcttcatc
     1861 actgacttcc tggacaatgg ctatgggcac tgtctcttag acaaaccaga ggctccattg
     1921 catctgcctg tgactttccc tggcaaggac tatgatgctg accgccagtg ccagctgacc
     1981 ttcgggcccg actcacgcca ttgtccacag ctgccgccgc cctgtgctgc cctctggtgc
     2041 tctggccacc tcaatggcca tgccatgtgc cagaccaaac actcgccctg ggccgatggc
     2101 acaccctgcg ggcccgcaca ggcctgcatg ggtggtcgct gcctccacat ggaccagctc
     2161 caggacttca atattccaca ggctggtggc tggggtcctt ggggaccatg gggtgactgc
     2221 tctcggacct gtgggggtgg tgtccagttc tcctcccgag actgcacgag gcctgtcccc
     2281 cggaatggtg gcaagtactg tgagggccgc cgtacccgct tccgctcctg caacactgag
     2341 gactgcccaa ctggctcagc cctgaccttc cgcgaggagc agtgtgctgc ctacaaccac
     2401 cgcaccgacc tcttcaagag cttcccaggg cccatggact gggttcctcg ctacacaggc
     2461 gtggcccccc aggaccagtg caaactcacc tgccaggccc aggcactggg ctactactat
     2521 gtgctggagc cacgggtggt agatgggacc ccctgttccc cggacagctc ctcggtctgt
     2581 gtccagggcc gatgcatcca tgctggctgt gatcgcatca ttggctccaa gaagaagttt
     2641 gacaagtgca tggtgtgcgg aggggacggt tctggttgca gcaagcagtc aggctccttc
     2701 aggaaattca ggtacggata caacaatgtg gtcactatcc ccgcgggggc cacccacatt
     2761 cttgtccggc agcagggaaa ccctggccac cggagcatct acttggccct gaagctgcca
     2821 gatggctcct atgccctcaa tggtgaatac acgctgatgc cctcccccac agatgtggta
     2881 ctgcctgggg cagtcagctt gcgctacagc ggggccactg cagcctcaga gacactgtca
     2941 ggccatgggc cactggccca gcctttgaca ctgcaagtcc tagtggctgg caacccccag
     3001 gacacacgcc tccgatacag cttcttcgtg ccccggccga ccccttcaac gccacgcccc
     3061 actccccagg actggctgca ccgaagagca cagattctgg agatccttcg gcggcgcccc
     3121 tgggcgggca ggaaactgaa ttctgcagtc gacggtaccc tggtgccgcg cggcagccat
     3181 atggctagct ggagccatcc gcagtttgaa aaaggtggtg gtagcggtgg tggttcaggt
     3241 ggtagtgcat ggtcacaccc tcagtttgag aaactggatc caccggtcgc caccatggtg
     3301 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     3361 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     3421 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     3481 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     3541 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     3601 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     3661 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     3721 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     3781 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     3841 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3901 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     3961 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     4021 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     4081 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     4141 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4201 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     4261 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4321 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     4381 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     4441 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     4501 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     4561 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     4621 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     4681 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     4741 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4801 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     4861 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4921 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4981 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5041 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5101 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     5161 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     5221 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5281 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5341 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5401 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     5461 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5521 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5581 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5641 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5701 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5761 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5821 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5881 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5941 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     6001 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     6061 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     6121 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     6181 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     6241 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     6301 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     6361 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     6421 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     6481 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6541 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6601 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6661 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6721 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6781 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6841 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6901 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6961 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     7021 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7081 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7141 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7201 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7261 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     7321 ctgtggataa ccgtattacc gccatgcat
//
