LOCUS       V047155                 7848 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047155
VERSION     V047155
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..2528
                     /label="Canx(NM_007597)"
                     /note="Canx(NM_007597)"
                     /gene="Canx"
     CDS             2535..2600
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2624..3212
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3278..3326
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3528..3956
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3970..4299
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4366..5160
                     /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    5336..5457
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5506..5522)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5530..5546
                     /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(5554..5584)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5599..5620
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5908..6496)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6667..7527)
                     /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(7528..7632)
                     /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 agggaagtgg ttactgtgtt
      781 tgctgctggt ccttggaact gcagctgttg aggctcatga tggacatgat gatgacgcga
      841 ttgatattga agatgatctt gatgatgtta ttgaagaggt agaagattca aaatctaaat
      901 cagatgccag cactcctcca tctccaaagg tcacctacaa agctccagtt ccaacagggg
      961 aggtttattt tgctgactcc tttgacagag ggtctctgtc agggtggatt ttatctaaag
     1021 ccaaaaaaga tgacactgat gatgaaattg ccaaatatga tggaaagtgg gaagtagatg
     1081 agatgaagga aacaaagctt ccaggggata aaggacttgt actgatgtct cgggccaagc
     1141 atcatgccat ctctgctaaa ctgaataagc ccttcctgtt tgataccaag cctctcattg
     1201 ttcagtatga ggttaatttt cagaatggaa tagaatgtgg tggtgcctat gtgaagctgc
     1261 tttccaagac ggcagagctc agcctggatc aattccacga caagactccc tatactatta
     1321 tgtttggtcc agataagtgt ggagaggact acaaactgca tttcatcttt cgacacaaaa
     1381 atcccaagac aggtgtatat gaagaaaaac atgctaagag gccagatgca gatctgaaga
     1441 cctatttcac tgacaagaaa acgcatcttt atacattaat cttgaatcca gacaatagtt
     1501 ttgaaatatt agttgaccag tctgttgtga acagtggaaa tctgctaaat gacatgactc
     1561 ctcctgtaaa cccttcacgt gaaattgaag acccagaaga ccggaagcct gaagattggg
     1621 atgaaaggcc caaaatagca gatccagatg ctgtcaagcc agatgactgg gatgaagacg
     1681 ccccttctaa gatcccagat gaagaggcca ccaagcctga aggctggcta gacgacgaac
     1741 ctgagtatat tccagaccct gatgcagaga agccagagga ttgggatgag gatatggacg
     1801 gagaatggga ggctcctcag attgccaacc ccaagtgtga gtcagcccct gggtgtggtg
     1861 tctggcagcg acctatgatt gacaacccca attataaggg caaatggaag cctccaatga
     1921 ttgacaaccc taactaccag ggaatctgga aaccaaggaa aataccaaat ccagatttct
     1981 ttgaagacct agaacctttt aagatgactc ctttcagtgc tattggtttg gagctctggt
     2041 ccatgacatc cgacatcttt tttgacaact ttatcattag tggtgaccga agagtagttg
     2101 atgattgggc caatgatggg tggggcctga agaaagctgc tgatggggct gctgagccag
     2161 gtgtagtgct gcagatgctg gaggcagctg aagagcgtcc atggctttgg gtggtctaca
     2221 ttttgactgt agctttgcca gtgttccttg tgatcctctt ctgctgttct ggaaagaaac
     2281 agtccaatgc tatggagtac aagaagacgg atgctcccca gccagatgtg aaggatgaag
     2341 aagggaagga agaagagaag aacaagaggg atgaagagga agaagaggag aagcttgaag
     2401 agaaacagaa gagtgatgct gaagaagatg gtgttactgg cagtcaagat gaggaagata
     2461 gcaagcctaa agcagaggag gatgaaattt tgaacagatc gccaagaaac agaaagccac
     2521 gaagagaggg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     2581 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     2641 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2701 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2761 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2821 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2881 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2941 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     3001 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     3061 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3121 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3181 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     3241 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     3301 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     3361 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     3421 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     3481 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     3541 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3601 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3661 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     3721 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3781 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3841 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     3901 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3961 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4021 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4081 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4141 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4201 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4261 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4321 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4381 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4441 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4501 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4561 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4621 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4681 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4741 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4801 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4861 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4921 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4981 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5041 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5101 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5161 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5221 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5281 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5341 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5401 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5461 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5521 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5581 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5641 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5701 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5761 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5821 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5881 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5941 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6001 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6061 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     6121 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6181 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6241 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6301 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6361 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6421 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6481 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6541 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6601 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6661 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6721 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6781 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6841 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6901 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6961 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7021 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7081 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7141 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7201 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7261 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7321 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7381 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7441 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7501 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7561 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7621 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     7681 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     7741 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     7801 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
