LOCUS       V018417                 7973 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018417
VERSION     V018417
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     166..182
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        343..722
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        723..926
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        952..970
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             1055..4144
                     /label="SLC12A3(NM_000339)"
                     /note="SLC12A3(NM_000339)"
                     /gene="SLC12A3"
     CDS             4151..4168
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     CDS             4187..4210
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(4230..4246)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    4257..4879
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(5028..5616)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    5945..5992
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(6224..7018)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     promoter        complement(7053..7410)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(7412..7516)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      7543..7971
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgcggc gcgccagatc tcaagcttgc caccatggca gaactgccca caacagagac
     1081 gcctggggac gccactttgt gcagcgggcg cttcaccatc agcacactgc tgagcagtga
     1141 tgagccctct ccaccagctg cctatgacag cagccacccc agccacctga cccacagcag
     1201 caccttctgc atgcgcacct ttggctacaa cacgatcgat gtggtgccca catatgagca
     1261 ctatgccaac agcacccagc ctggtgagcc ccggaaggtc cggcccacac tggctgacct
     1321 gcactccttc ctcaagcagg aaggcagaca cctgcatgcc ctggcctttg acagccggcc
     1381 cagccacgag atgactgatg ggctggtgga gggcgaggca ggcaccagca gcgagaagaa
     1441 ccccgaggag ccagtgcgct tcggctgggt caagggggtg atgattcgtt gcatgctcaa
     1501 catttggggc gtgatcctct acctgcggct gccctggatt acggcccagg caggcatcgt
     1561 cctgacctgg atcatcatcc tgctgtcggt cacggtgacc tccatcacag gcctctccat
     1621 ctcagccatc tccaccaatg gcaaggtcaa gtcaggtggc acctacttcc tcatctcccg
     1681 gagtctgggc ccagagcttg ggggctccat cggcctcatt ttcgctttcg ccaatgccgt
     1741 gggtgtggcc atgcacacgg tgggctttgc agagaccgtg cgggacctgc tccaggagta
     1801 tggggcaccc atcgtggacc ccattaacga catccgcatc attgccgtgg tctcggtcac
     1861 tgtgctgctg gccatctccc tggctggcat ggagtgggag tccaaggccc aggtgctgtt
     1921 cttccttgtc atcatggtct cctttgccaa ctatttagtg gggacgctga tccccccatc
     1981 tgaggacaag gcctccaaag gcttcttcag ctaccgggcg gacatttttg tccagaactt
     2041 ggtgcctgac tggcggggtc cagatggcac cttcttcgga atgttctcca tcttcttccc
     2101 ctcggccaca ggcatcctgg caggggccaa catatctggt gacctcaagg accctgctat
     2161 agccatcccc aaggggaccc tcatggccat tttctggacg accatttcct acctggccat
     2221 ctcagccacc attggctcct gcgtggtgcg tgatgcctct ggggtcctga atgacacagt
     2281 gacccctggc tggggtgcct gcgaggggct ggcctgcagc tatggctgga acttcaccga
     2341 gtgcacccag cagcacagct gccactacgg cctcatcaac tattaccaga ccatgagcat
     2401 ggtgtcaggc ttcgcgcccc tgatcacggc tggcatcttc ggggccaccc tctcctctgc
     2461 cctggcctgc cttgtctctg ctgccaaagt cttccagtgc ctttgcgagg accagctgta
     2521 cccactgatc ggcttcttcg gcaaaggcta tggcaagaac aaggagcccg tgcgtggcta
     2581 cctgctggcc tacgccatcg ctgtggcctt catcatcatc gctgagctca acaccatagc
     2641 ccccatcatt tccaacttct tcctctgctc ctatgccctc atcaacttca gctgcttcca
     2701 cgcctccatc accaactcgc ctgggtggag accttcattc caatactaca acaagtgggc
     2761 ggcgctgttt ggggctatca tctccgtggt catcatgttc ctcctcacct ggtgggcggc
     2821 cctcatcgcc attggcgtgg tgctcttcct cctgctctat gtcatctaca agaagccaga
     2881 ggtaaattgg ggctcctcgg tacaggctgg ctcctacaac ctggccctca gctactcggt
     2941 gggcctcaat gaggtggaag accacatcaa gaactaccgc ccccagtgcc tggtgctcac
     3001 ggggcccccc aacttccgcc cggccctggt ggactttgtg ggcaccttca cccggaacct
     3061 cagcctgatg atctgtggcc acgtgctcat cggaccccac aagcagagga tgcctgagct
     3121 ccagctcatc gccaacgggc acaccaagtg gctgaacaag aggaagatca aggccttcta
     3181 ctcggatgtc attgccgagg acctccgcag aggcgtccag atcctcatgc aggccgcagg
     3241 tctcgggaga atgaagccca acattctggt ggttgggttc aagaagaact ggcagtcggc
     3301 tcacccggcc acagtggaag actacattgg catcctccat gatgcctttg atttcaacta
     3361 tggcgtgtgt gtcatgagga tgcgggaggg actcaacgtg tccaagatga tgcaggcgca
     3421 cattaacccc gtgtttgacc cagcggagga cgggaaggaa gccagcgcca gaggtgccag
     3481 gccatcagtc tctggcgctt tggaccccaa ggccctggtg aaggaggagc aggccaccac
     3541 catcttccag tcggagcagg gcaagaagac catagacatc tactggctct ttgacgatgg
     3601 aggcctcacc ctcctcattc cctatctcct tggccgcaag aggaggtgga gcaaatgcaa
     3661 gatccgtgtg ttcgtaggcg gccagattaa caggatggac caggagagaa aggcgatcat
     3721 ttctctgctg agcaagttcc gactgggatt ccatgaagtc cacatcctcc ctgacatcaa
     3781 ccagaaccct cgggctgagc acaccaagag gtttgaggac atgattgcac ccttccgtct
     3841 gaatgatggc ttcaaggatg aggccactgt caacgagatg cggcgggact gcccctggaa
     3901 gatctcagat gaggagatta cgaagaacag agtcaagtcc cttcggcagg tgaggctgaa
     3961 tgagattgtg ctggattact cccgagacgc tgctctcatc gtcatcactt tgcccatagg
     4021 gaggaagggg aagtgcccca gctcgctgta catggcctgg ctggagaccc tgtcccagga
     4081 cctcagacct ccagtcatcc tgatccgagg aaaccaggaa aacgtgctca ccttttactg
     4141 ccagctcgag catcatcacc atcaccatgc agcaaatgat atcctggatt acaaggatga
     4201 cgacgataag gtttaaacgg ccggccgcgg tcatagctgt ttcctgaaca gatcccgggt
     4261 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     4321 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     4381 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     4441 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     4501 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     4561 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     4621 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     4681 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     4741 taaaataact ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc
     4801 aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc
     4861 agtagatgcc tgttgaattg ggtacgcggc cagcggcgag cggtatcagc tcactcaaag
     4921 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4981 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5041 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5101 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5161 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5221 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5281 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5341 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5401 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5461 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5521 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5581 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5641 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5701 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5761 atatatgagt aacctgaggc tatggcaggg cctgccgccc cgacgttggc tgcgagccct
     5821 gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg
     5881 ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt
     5941 ttatgaacaa acgacccaac accgtgcgtt ttattctgtc tttttattgc cgtcatagcg
     6001 cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccct agggtgggcg
     6061 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     6121 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
     6181 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     6241 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     6301 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     6361 cctgatagcg atccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     6421 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     6481 cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     6541 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     6601 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     6661 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     6721 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     6781 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca
     6841 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     6901 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     6961 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     7021 gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa agcctaggcc tccaaaaaag
     7081 cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata
     7141 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     7201 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
     7261 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
     7321 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
     7381 acattccaca gctggttctt tccgcctcag gactcttcct ttttcaatat tattgaagca
     7441 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7501 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     7561 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     7621 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     7681 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     7741 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     7801 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     7861 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     7921 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat ttt
//
