LOCUS       V030556                 9166 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030556
VERSION     V030556
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..4659
                     /label="WNK4(NM_032387)"
                     /note="WNK4(NM_032387)"
                     /gene="WNK4"
     CDS             4660..4686
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    4733..4957
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5003..5431
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5445..5774
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5841..6635
                     /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    6809..6930
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6979..6995)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7003..7019
                     /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(7027..7057)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7072..7093
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7381..7966)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8137..8997)
                     /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(8998..9102)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atgttggcat ccccggccac ggagaccacc
      961 gtcctcatgt cccagactga ggccgacctg gccctgcggc ccccgcctcc tcttggcacc
     1021 gcggggcagc cccgcctcgg gccccctcct cgccgagcgc gccgcttctc cgggaaggct
     1081 gagccccggc cgcgctcttc tcgtctcagc cgccgtagct cagtcgactt ggggctgctg
     1141 agctcttggt ccctgccagc ctcacccgct ccggaccccc ccgatcctcc ggactccgct
     1201 ggtcctggcc ccgcgaggag cccaccgcct agctccaaag aaccccccga gggcacgtgg
     1261 accgagggag cccctgtgaa ggctgcggaa gactccgcgc gtcccgagct cccggactct
     1321 gcagtgggcc cggggtccag ggagccgcta agggtccctg aagctgtggc cctagagcgg
     1381 cggcgggagc aggaagaaaa ggaggacatg gagacccagg ctgtggcaac gtcccccgat
     1441 ggccgatacc tcaagtttga catcgagatt ggacgtggct ccttcaagac ggtgtatcga
     1501 gggctagaca ccgacaccac agtggaggtg gcctggtgtg agctgcagac tcggaaactg
     1561 tctagagctg agcggcagcg cttctcagag gaggtggaga tgctcaaggg gctgcagcac
     1621 cccaacatcg tccgcttcta tgattcgtgg aagtcggtgc tgaggggcca ggtttgcatc
     1681 gtgctggtca ccgaactcat gacctcgggc acgctcaaga cgtacctgag gcggttccgg
     1741 gagatgaagc cgcgggtcct tcagcgctgg agccgccaaa tcctgcgggg acttcatttc
     1801 ctacactccc gggttcctcc catcctgcac cgggatctca agtgcgacaa tgtctttatc
     1861 acgggaccta ctggctctgt caaaatcggg gacctgggcc tggccacgct caagcgcgcc
     1921 tcctttgcca agagtgtcat cgggaccccg gaattcatgg cccccgagat gtacgaggaa
     1981 aagtacgatg aggccgtgga cgtgtacgcg ttcggcatgt gcatgctgga gatggccacc
     2041 tctgagtacc cgtactccga gtgccagaat gccgcgcaaa tctaccgcaa ggtcacttcg
     2101 ggcagaaagc cgaacagctt ccacaaggtg aagatacccg aggtgaagga gatcattgaa
     2161 ggctgcatcc gcacggataa gaacgagagg ttcaccatcc aggacctcct ggcccacgcc
     2221 ttcttccgcg aggagcgcgg tgtgcacgtg gaactagcgg aggaggacga cggcgagaag
     2281 ccgggcctca agctctggct gcgcatggag gacgcgcggc gcggggggcg cccacgggac
     2341 aaccaggcca tcgagttcct gttccagctg ggccgggacg cggccgagga ggtggcacag
     2401 gagatggtgg ctctgggctt ggtctgtgaa gccgattacc agccagtggc ccgtgcagta
     2461 cgtgaacggg ttgctgccat ccagcgaaag cgtgagaagc tgcgtaaagc aagggaattg
     2521 gaggcactcc caccagagcc aggacctcca ccagcaactg tgcccatggc ccccggtccc
     2581 cccagtgtct tcccccctga gcctgaggag ccagaggcag accagcacca gcccttcctt
     2641 ttccgccacg ccagctactc atctaccact tcggattgcg agactgatgg ctacctcagc
     2701 tcctccggct tcctggatgc ctcagaccct gcccttcagc cccctggggg ggtgccatcc
     2761 agcctggctg agtcccatct ctgcctgccc tcggcttttg ccctatccat tccacgttct
     2821 ggccctggaa gtgacttttc ccccggggac agctatgcct cagatgcagc ttcaggcctt
     2881 agcgatgtgg gagaagggat gggacaaatg aggagacccc cagggaggaa tctccggcgc
     2941 agaccccgat cccggctgcg ggtcactagt gtctcagacc agaatgacag agtggttgag
     3001 tgccagctac agacccataa cagcaagatg gtgaccttcc gatttgatct ggatggggac
     3061 agcccggaag agattgcagc tgccatggta tataacgagt tcattctgcc ttcggagcga
     3121 gatggatttc tcagacggat tcgggagatt atccagcgag tggagaccct gttgaagaga
     3181 gacactggcc ccatggaggc tgctgaagac accctaagcc cccaggagga gccagcacca
     3241 ttacctgccc tgcccgtccc cctcccagac ccatccaatg aagagctcca gagcagcacc
     3301 tccctggagc acaggagctg gacagccttc tccacctcct catcttctcc tggaactcct
     3361 ttgtctcctg gaaacccatt ttcccctgga acccccattt ccccaggtcc catcttcccc
     3421 atcacttctc ccccatgtca tcccagcccc tccccattct cccccatttc ttcccaggtc
     3481 tcctcaaatc cctctccaca ccccaccagc tctccacttc cattctcctc cagcacaccc
     3541 gagtttccgg tcccactctc tcagtgtccc tggagttctc tccccacgac ttctccacct
     3601 acgttctctc ccacttgttc tcaggtcact cttagttccc ctttctttcc tccgtgcccc
     3661 tccacttctt ccttcccctc caccacagca gcccctctcc tttctctggc tagtgccttc
     3721 tcactggctg tgatgactgt ggcccagtcc ctgctgtccc cctcacctgg gctcctttcc
     3781 cagtctcctc cagcccctcc tagtcccctc cctagcctgc cccttccccc tcccgttgct
     3841 cctggtggcc aggaaagccc ttcaccccac acagctgagg tggagagtga ggcctcacca
     3901 cctcctgctc ggcccctccc aggggaagcc aggctggcgc ccatctctga agagggaaag
     3961 ccgcagcttg ttgggcgttt ccaagtgact tcatccaagg aaccggctga gcctcttccc
     4021 ttgcagccaa catcccccac tctctctggt tctccaaaac cttcaacccc tcagctcact
     4081 tcagagagct cagatacaga ggacagtgct ggaggcgggc cagagaccag ggaagctctg
     4141 gctgagagcg accgtgcagc tgagggtctg ggggctggag ttgaggagga aggagatgat
     4201 gggaaggaac cccaagttgg gggcagcccc caacccctga gccatcccag cccagtgtgg
     4261 atgaactact cctacagcag cctgtgtttg agcagcgagg agtcagaaag cagtggggaa
     4321 gatgaggagt tctgggctga gctgcagagt cttcggcaga agcacttgtc agaggtggaa
     4381 acactacaga cactacagaa aaaagaaatt gaagatttgt acagccggct ggggaagcag
     4441 cccccaccgg gtattgtggc cccagctgct atgctgtcca gccgccagcg ccgcctctcc
     4501 aagggcagct tccccacctc ccgccgcaac agcctacagc gctctgagcc cccaggccct
     4561 ggcatcatgc gaaggaactc tctgagtggc agcagcaccg gctcccagga gcagcgggca
     4621 agcaaggggg tgacattcgc cggggatgtt ggcaggatgt acccatacga tgtgccagat
     4681 tacgcatagc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     4741 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     4801 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     4861 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     4921 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     4981 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5041 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     5101 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     5161 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     5221 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     5281 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5341 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5401 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     5461 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5521 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5581 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5641 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5701 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5761 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     5821 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5881 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5941 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     6001 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     6061 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     6121 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     6181 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     6241 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     6301 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     6361 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     6421 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     6481 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     6541 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     6601 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     6661 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     6721 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6781 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     6841 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6901 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     6961 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     7021 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     7081 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     7141 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     7201 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     7261 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7321 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7381 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7441 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7501 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7561 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7621 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7681 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7741 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7801 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     7861 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     7921 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7981 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     8041 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8101 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     8161 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     8221 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8281 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8341 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8401 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8461 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8521 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8581 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8641 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8701 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8761 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8821 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8881 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8941 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     9001 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9061 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9121 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggt
//
