LOCUS       V032238                 9456 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032238
VERSION     V032238
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             919..4269
                     /label="Adamts6(NM_001081020)"
                     /note="Adamts6(NM_001081020)"
                     /gene="Adamts6"
     CDS             4306..5025
                     /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    5056..5280
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5326..5754
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5768..6097
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6164..6958
                     /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    7132..7253
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7302..7318)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7326..7342
                     /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(7350..7380)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7395..7416
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7704..8289)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8460..9320)
                     /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(9321..9425)
                     /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 aagccaccat ggaaatttcg tggaaaacgt tgacttggat tctgagcctc
      961 atcatggcct catcggaatt ttatagtgac cacagacttt catatagttc tcaagaggaa
     1021 ttcctgactt atcttgaaca ctaccagcta actattccaa taagagttga ccaaaatgga
     1081 gcttttctca gctttactgt gaaaaatgat aaacactcaa gaagaagacg gagtatggac
     1141 cttcttgatc cacagcaggc agtgtctaaa ttatttttta aactttcagc ctatggcaag
     1201 cactttcacc taaacttgac tctcaataca aattttgtgt ccaaacattt tacagtagag
     1261 tattggggga aagatggacc ccagtggaaa catgattttt tagacaactg tcactacaca
     1321 ggatacttgc aagatcaaca tagtacaact aaagtggctt taagcaactg tgttggattg
     1381 catggtatta ttgctacaga agatgaagaa tattttattg agcctctaaa gaatactaca
     1441 gaggattcca aacattttag ttatgaaaat ggccatcctc atgttattta caaaaagtct
     1501 actcttcagc aacaacatat ctatgatcat tctcattgtg gggtttcaga tctcacagga
     1561 agtggcaaac cttggtggct gaataagaca tcatcttttc cttctttacc accaattaat
     1621 gacacacata ttcatcacag aaagagaaga tctgtgagca ctgagagatt tgtggagact
     1681 ttagtagtgg cagacaaaat gatggtgggc taccatggcc gtaaagacat tgaacattat
     1741 attttaagtg tgatgaatat tgttgccaaa ctttatcgtg actccagtct aggaaacgtt
     1801 gtgaatatta tagtggctcg cttaattgtt ctcactgaag atcagccaaa cttggagata
     1861 aaccatcatg cagacaagtc cctcgatagc ttctgtaaat ggcagaaatc cattctctcc
     1921 caccaaagtg atggaaacac cattccagaa aatgggattg cccaccacga taatgcagtt
     1981 cttattacta gatatgatat ctgcacgtat aaaaacaagc cctgtggaac actgggctta
     2041 gcctctgtgg ctggaatgtg tgagcctgaa aggagctgca gcattaatga agacattggc
     2101 ctaggctcgg cttttaccat cgcacatgag attggtcaca attttggtat gaaccatgat
     2161 ggaattggaa attcttgtgg gacgaaaggt catgaggcag caaaacttat ggcagctcat
     2221 attactgcaa ataccaatcc cttctcctgg tctgcctgca gcagagacta catcaccagc
     2281 tttctagatt caggccgtgg tacttgcctt gataatgagc ctccaaagcg tgactttctt
     2341 tatccagctg tggccccagg tcaggtgtat gatgctgatg aacaatgtcg tttccagtat
     2401 ggagcaacat cccgccaatg caaatatggg gaagtgtgta gagagctctg gtgcctcagc
     2461 aaaagcaacc gctgtgtcac caacagtatt ccagctgctg aggggacact gtgtcagact
     2521 gggaatattg agaaagggtg gtgttatcag ggagattgtg ttccttttgg cacatggccc
     2581 cagagtattg atgggggctg gggttcctgg tcactgtggg gcgagtgcag taggacctgc
     2641 ggaggaggtg tctcctcgtc tctaagacac tgtgacagtc cagcaccttc aggaggtggg
     2701 aaatattgcc ttggtgaaag gaaacgctac cgctcttgca acacagatcc atgccctttg
     2761 ggttcccgag attttcgaga aaaacagtgt gcagactttg acagtatgcc tttccgggga
     2821 aagcattata actggaaacc ctatactgga ggtggggtaa aaccttgtgc attaaattgc
     2881 ctggctgaag gctataattt ctacacagag cgagctcctg ctgtgataga tgggacacag
     2941 tgcaatgcag actcactgga tatctgtatc aatggagaat gcaagcacgt aggttgtgat
     3001 aatattttgg gatctgatgc tagggaagat aggtgccgag tctgtggtgg ggatggaagc
     3061 acctgtgatg ccattgaagg gttcttcaat gactcattgc ccagaggagg ttatatggaa
     3121 gtggttcaga taccaagagg ctctgttcat attgaagtaa gagaagtttc catgtcaaaa
     3181 aattacattg ctttaaaatc tgaaggggat gattactaca ttaatggtgc ctggactatt
     3241 gactggccta ggaaatttga tgtttcgggg acagcttttc actacaagag accgacagat
     3301 gaaccagaat ccttggaagc tctaggaccc acctctgaga acctcattgt aatggttcta
     3361 ctccaagaac agaatttggg aattaggtac aagttcaatg ttcccatcgt tcgtactggc
     3421 agtggagata atgaagttgg cttcatgtgg acttatcagc cctgggcgga gtgctctgct
     3481 acttgtgctg gaggtgtcca aagacaagaa gtggtctgta aaagattgga tgacaactcc
     3541 attgtccaga acaatttctg tgatcctgac agtaagccac ctgaaaatca aagagcctgc
     3601 aacactgagc cttgcccacc tgaatggttc attggagact ggttggagtg cagcaagact
     3661 tgtgatggtg ggatgcgcac aagggccgtg ctctgcatca ggaaagttgg accttctgaa
     3721 gaggaaacac tggactacgg cgactgctta acacaccggc ctgtggagaa agagtcctgc
     3781 aacaaccagt catgtccacc tcagtgggta gctttggact ggtccgaatg tactcctaaa
     3841 tgtggttcag gattcaagca tcggattgtt ctgtgcaaga gcagtgacct ttctaaaaca
     3901 ttcccagctg cacaatgtcc agaggagagc aagcctcctg ctcgcatccg ctgcagtctg
     3961 ggccgctgcc ctcctccccg ctgggtgaca ggagactggg gccagtgctc tgctcagtgt
     4021 ggtcttggac aacaaatgag aactgtccag tgcctttcct atactggaca ggcgtctgtt
     4081 gactgtccag aaactgtccg gcctccatcc atgcagcagt gtgacagcaa atgtgatagc
     4141 acccctctct ccagtactga agagtgcaaa gatgtgaaca aagtggctta ttgcccactg
     4201 gtgctgaagt tcaagttctg cagtcgagca tacttcagac agatgtgttg taagacctgc
     4261 caaggacact cgagtggcgg tggaggaagc ggcggaggtg gatctatggt gagcaagggc
     4321 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     4381 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     4441 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     4501 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     4561 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     4621 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     4681 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     4741 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     4801 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     4861 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     4921 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     4981 accgccgccg ggatcactct cggcatggac gagctgtaca agtagggccc gtttaaaccc
     5041 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     5101 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     5161 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     5221 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     5281 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     5341 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     5401 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     5461 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     5521 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     5581 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5641 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     5701 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     5761 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5821 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     5881 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     5941 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6001 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     6061 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     6121 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     6181 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     6241 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     6301 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     6361 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     6421 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     6481 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     6541 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     6601 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     6661 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     6721 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     6781 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     6841 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     6901 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     6961 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     7021 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     7081 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     7141 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     7201 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     7261 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     7321 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     7381 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     7441 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     7501 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     7561 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7621 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7681 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7741 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7801 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7861 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7921 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7981 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     8041 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     8101 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     8161 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     8221 caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     8281 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     8341 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     8401 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     8461 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     8521 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     8581 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     8641 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     8701 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     8761 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     8821 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     8881 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     8941 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     9001 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     9061 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     9121 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     9181 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     9241 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     9301 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     9361 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     9421 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
