LOCUS       V041086                 7200 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041086
VERSION     V041086
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..1880
                     /label="Ciart(NM_001033302)"
                     /note="Ciart(NM_001033302)"
                     /gene="Ciart"
     CDS             1887..1952
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1976..2564
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2630..2678
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2880..3308
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3322..3651
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3718..4512
                     /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    4688..4809
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4858..4874)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4882..4898
                     /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(4906..4936)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4951..4972
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5260..5848)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6019..6879)
                     /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(6880..6984)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgga ttctccatct agtgtttctt
      781 cttattcctc ctcctccctc tccccatcat tttccacgtc ctctgtgaac agtgacttta
      841 gcttcccttc tgataacgag agggagggca agggcaccca tgagctcagg ccagacactg
      901 tgggtcagag aggaggttcc aggcccagtc caggccctat ccgctgcaga catcgaccca
      961 gggtttctag taaccaacat acagcacctc atctggaaca gcaaggatct gaagtcaaga
     1021 gatcaagaga cggtgaacag gagacgagtc tgaacaccca aggatgtact acagagggag
     1081 atctgctctt tgctcaaaag tgtaaagaac tccaagggtt catacggccc ctcacagacc
     1141 tactgaacgg actcaagatg ggtcgctttg acagaggatt aagtagcttc caacagagtg
     1201 tggccatgga ccggatccag cgcatagtgg gtgttttaca gaagcctcag atgggagaac
     1261 gttatctagg aaccttgctg caggtggaag ggatgttaaa gacttggttt cctcacatag
     1321 ctgcccagaa gtcatcctca ggaggtagcc ggcaccagat aagcaagcac tttcccagcc
     1381 accacggtga ccccggggct gcttctcctg cacctctcct agagaagatg ggccaaacac
     1441 agctgggaca tttagttttg aaaccaaagc agccttggca tctcactgga tggccagcta
     1501 tgaacctcac ctggattcat agcactccaa tttgtaaccc tcccctcagc tcccaaggct
     1561 ccgcctctgg ccacagcccc ataggcactg gagccagcat tggtgtcatc cttgtcctcc
     1621 agaaaggagg acagcccttc acccattctg ccccgggcac tccggtccca ccaactccac
     1681 tgtctcctgt cgtccctggt gatctgaaga aactgcctgg cgaggagcct cgttgccaca
     1741 gtttgccagt aacactgcca tcagactgga gttgcatcct atgtccgcct gtcctgccta
     1801 ccacagacag agagatgact aaaggacacc cagagccaca gatgactagc catcccccag
     1861 ttgctccgga tcctcagccc ggatccgact acaaagacca tgacggtgat tataaagatc
     1921 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     1981 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2041 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2101 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2161 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     2221 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     2281 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     2341 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     2401 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     2461 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     2521 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     2581 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     2641 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     2701 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     2761 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     2821 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     2881 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2941 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3001 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     3061 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3121 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3181 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3241 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3301 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3361 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3421 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3481 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3541 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3601 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3661 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     3721 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3781 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3841 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3901 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3961 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4021 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4081 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4141 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4201 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4261 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4321 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4381 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4441 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4501 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     4561 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4621 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4681 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4741 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4801 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4861 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4921 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4981 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5041 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5101 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5161 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5221 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5281 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5341 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5401 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     5461 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5521 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5581 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5641 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5701 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5761 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5821 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5881 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5941 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6001 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6061 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6121 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6181 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6241 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6301 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6361 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6421 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6481 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6541 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6601 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6661 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6721 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6781 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6841 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6901 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6961 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     7021 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     7081 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     7141 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
