LOCUS       V018828                 7580 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018828
VERSION     V018828
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             671..2260
                     /label="CATSPER2(NM_172095)"
                     /note="CATSPER2(NM_172095)"
                     /gene="CATSPER2"
     CDS             2267..2332
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2356..2944
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3010..3058
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3260..3688
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3702..4031
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4098..4892
                     /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    5068..5189
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5238..5254)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5262..5278
                     /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(5286..5316)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5331..5352
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5640..6228)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6399..7259)
                     /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(7260..7364)
                     /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 taccgccacc atggccgctt accaacaaga agagcagatg cagcttcccc gagctgatgc
      721 cattcgttca cgtctcatcg atactttctc tctcattgag catttgcaag gcttgagcca
      781 agctgtgccg cggcacacta tcagggagtt acttgatcct tcccgccaga agaaacttgt
      841 attgggagat caacaccagc tagtgcgttt ctctataaag cctcagcgta tagaacagat
      901 ttcacatgcc cagaggctgt tgagcaggct tcatgtgcgc tgcagtcaga ggccacctct
      961 ttctttgtgg gccggatggg tccttgagtg tcctctcttc aaaaacttca tcatcttcct
     1021 ggtctttttg aatacgatca tattgatggt tgaaatagaa ttgctggaat ccacaaatac
     1081 caaactatgg ccattgaagc tgaccttgga ggtggcagct tggtttatct tgcttatttt
     1141 catcctggag atccttctta agtggctatc caacttttct gttttctgga agagtgcctg
     1201 gaatgtcttt gactttgttg ttaccatgtt gtccctgctt cccgaggttg tggtattggt
     1261 aggggtaaca ggccaatcgg tgtggcttca gcttctgagg atctgccggg tgctgaggtc
     1321 tctcaaactc cttgcacaat tccgtcaaat tcaaattatt attttggtcc tggtcagggc
     1381 cctcaagagc atgaccttcc tcttgatgtt gctgctcatc ttcttctaca tttttgctgt
     1441 gactggtgtc tacgtcttct cagagtacac ccgttcacct cgtcaggacc tggagtacca
     1501 tgtgttcttc tcggacctcc cgaattccct ggtaacagtg ttcattctct tcaccttgga
     1561 tcattggtat gcactgcttc aggacgtctg gaaggtgcct gaagtcagtc gcatcttcag
     1621 cagcatctat ttcatccttt ggttgttgct tggctccatt atctttcgaa gtatcatagt
     1681 agccatgatg gttactaact ttcagaatat caggaaagag ctgaatgagg agatggcgcg
     1741 tcgggaggtt cagctcaaag ctgacatgtt caagcggcag atcatccaga ggagaaaaaa
     1801 catgtcacat gaagcactga cgtcaagcca tagcaaaata gaggacagtt caagaggagc
     1861 tagtcaacaa agggaaagtt tggacttatc agaagtgtct gaagtagagt ctaattatgg
     1921 tgccactgaa gaggatttaa taacatctgc atcaaaaaca gaagagacct tgtcaaaaaa
     1981 gagagagtac cagtcttcct cctgtgtctc ctccacatcc tcttcctatt cttcctcttc
     2041 tgaatccaga ttttctgaat ctattggtcg tttggactgg gagactcttg tgcacgaaaa
     2101 tctgcccggg ctaatggaaa tggatcagga tgaccgtgtt tggcccagag actcactctt
     2161 ccgatatttt gagttgctag aaaagcttca gtataaccta gaggaacgta agaagttaca
     2221 agagtttgca gtgcaggcac tgatgaactt ggaagacaag ctcgaggact acaaagacca
     2281 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg
     2341 gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2401 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2461 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2521 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2581 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2641 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2701 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     2761 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     2821 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     2881 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     2941 ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     3001 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat
     3061 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     3121 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     3181 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg
     3241 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3301 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3361 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc
     3421 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3481 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3541 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3601 gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag
     3661 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3721 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3781 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3841 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3901 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3961 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4021 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4081 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4141 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4201 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4261 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4321 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4381 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4441 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4501 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4561 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4621 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4681 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4741 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4801 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4861 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc
     4921 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4981 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5041 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5101 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5161 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5221 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5281 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5341 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5401 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5461 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5521 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5581 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5641 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5701 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5761 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5821 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5881 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5941 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6001 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6061 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6121 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6181 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6241 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6301 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6361 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6421 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6481 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6541 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6601 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6661 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6721 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6781 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6841 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6901 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6961 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7021 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7081 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7141 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7201 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7261 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7321 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7381 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta
     7441 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     7501 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     7561 gcatgaagaa tctgcttagg
//
