LOCUS       V034378                 7778 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034378
VERSION     V034378
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"
     CDS             683..2458
                     /label="CANX(NM_001746)"
                     /note="CANX(NM_001746)"
                     /gene="CANX"
     CDS             2465..2530
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2554..3142
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3208..3256
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3458..3886
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3900..4229
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4296..5090
                     /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    5266..5387
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5436..5452)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5460..5476
                     /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(5484..5514)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5529..5550
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5838..6426)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6597..7457)
                     /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(7458..7562)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggaagg gaagtggttg ctgtgtatgt tactggtgct
      721 tggaactgct attgttgagg ctcatgatgg acatgatgat gatgtgattg atattgagga
      781 tgaccttgac gatgtcattg aagaggtaga agactcaaaa ccagatacca ctgctcctcc
      841 ttcatctccc aaggttactt acaaagctcc agttccaaca ggggaagtat attttgctga
      901 ttcttttgac agaggaactc tgtcagggtg gattttatcc aaagccaaga aagacgatac
      961 cgatgatgaa attgccaaat atgatggaaa gtgggaggta gaggaaatga aggagtcaaa
     1021 gcttccaggt gataaaggac ttgtgttgat gtctcgggcc aagcatcatg ccatctctgc
     1081 taaactgaac aagcccttcc tgtttgacac caagcctctc attgttcagt atgaggttaa
     1141 tttccaaaat ggaatagaat gtggtggtgc ctatgtgaaa ctgctttcta aaacaccaga
     1201 actcaacctg gatcagttcc atgacaagac cccttatacg attatgtttg gtccagataa
     1261 atgtggagag gactataaac tgcacttcat cttccgacac aaaaacccca aaacgggtat
     1321 ctatgaagaa aaacatgcta agaggccaga tgcagatctg aagacctatt ttactgataa
     1381 gaaaacacat ctttacacac taatcttgaa tccagataat agttttgaaa tactggttga
     1441 ccaatctgtg gtgaatagtg gaaatctgct caatgacatg actcctcctg taaatccttc
     1501 acgtgaaatt gaggacccag aagaccggaa gcccgaggat tgggatgaaa gaccaaaaat
     1561 cccagatcca gaagctgtca agccagatga ctgggatgaa gatgcccctg ctaagattcc
     1621 agatgaagag gccacaaaac ccgaaggctg gttagatgat gagcctgagt acgtacctga
     1681 tccagacgca gagaaacctg aggattggga tgaagacatg gatggagaat gggaggctcc
     1741 tcagattgcc aaccctagat gtgagtcagc tcctggatgt ggtgtctggc agcgacctgt
     1801 gattgacaac cccaattata aaggcaaatg gaagcctcct atgattgaca atcccagtta
     1861 ccagggaatc tggaaaccca ggaaaatacc aaatccagat ttctttgaag atctggaacc
     1921 tttcagaatg actcctttta gtgctattgg tttggagctg tggtccatga cctctgacat
     1981 tttttttgac aactttatca tttgtgctga tcgaagaata gttgatgatt gggccaatga
     2041 tggatggggc ctgaagaaag ctgctgatgg ggctgctgag ccaggcgttg tggggcagat
     2101 gatcgaggca gctgaagagc gcccgtggct gtgggtagtc tatattctaa ctgtagccct
     2161 tcctgtgttc ctggttatcc tcttctgctg ttctggaaag aaacagacca gtggtatgga
     2221 gtataagaaa actgatgcac ctcaaccgga tgtgaaggaa gaggaagaag agaaggaaga
     2281 ggaaaaggac aagggagatg aggaggagga aggagaagag aaacttgaag agaaacagaa
     2341 aagtgatgct gaagaagatg gtggcactgt cagtcaagag gaggaagaca gaaaacctaa
     2401 agcagaggag gatgaaattt tgaacagatc accaagaaac agaaagccac gaagagagct
     2461 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     2521 cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt
     2581 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2641 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2701 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2761 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2821 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2881 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2941 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     3001 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3061 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3121 tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga
     3181 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     3241 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     3301 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     3361 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     3421 gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3481 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3541 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3601 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3661 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3721 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3781 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     3841 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3901 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3961 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4021 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4081 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4141 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4201 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4261 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4321 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4381 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4441 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     4501 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4561 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4621 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4681 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4741 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4801 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     4861 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4921 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4981 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5041 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5101 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5161 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5221 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5281 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5341 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5401 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5461 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5521 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5581 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5641 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5701 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5761 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5821 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5881 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5941 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6001 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     6061 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6121 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6181 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6241 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6301 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6361 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6421 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6481 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6541 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6601 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6661 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6721 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6781 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6841 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6901 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6961 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7021 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7081 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7141 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7201 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7261 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7321 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7381 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7441 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7501 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7561 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     7621 tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     7681 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     7741 aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
