LOCUS       V019030                 7505 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019030
VERSION     V019030
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..2185
                     /label="KCNJ3(NM_002239)"
                     /note="KCNJ3(NM_002239)"
                     /gene="KCNJ3"
     CDS             2192..2257
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2281..2869
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2935..2983
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3185..3613
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3627..3956
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4023..4817
                     /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    4993..5114
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5163..5179)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5187..5203
                     /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(5211..5241)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5256..5277
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5565..6153)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6324..7184)
                     /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(7185..7289)
                     /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 ccatgtctgc actccgaagg aaatttgggg acgattatca
      721 ggtagtgacc acatcgtcca gcggctcggg cttgcagccc caggggccag gccaggaccc
      781 tcagcagcag cttgtgccca agaagaagcg gcagcggttc gtggacaaga acggccggtg
      841 caatgtacag cacggcaacc tgggcagcga gacaagccgc tacctctcgg acctcttcac
      901 cacgctggtg gacctcaagt ggcgctggaa cctcttcatc ttcattctca cctacaccgt
      961 ggcctggctt ttcatggcgt ccatgtggtg ggtgatcgcc tacactcggg gcgacctgaa
     1021 caaagcccac gtcggtaact acacgccttg cgtggccaat gtctataact tcccttctgc
     1081 cttcctcttc ttcatcgaga cggaggccac catcggctat ggctaccgat acatcacaga
     1141 caagtgcccc gagggcatca tcctcttcct cttccagtcc atcctgggct ccatcgtgga
     1201 cgccttcctc atcggctgca tgttcatcaa gatgtcccag cccaagaagc gcgccgagac
     1261 cctcatgttc agcgagcacg cggtgatctc catgagggac ggaaaactca cgcttatgtt
     1321 ccgggtgggc aacctgcgca acagccacat ggtctccgcg cagattcgct gcaagctgct
     1381 caaatctcgg cagacacctg agggtgagtt ccttcccctt gaccaacttg aactggatgt
     1441 aggttttagt acaggggcag atcaactttt tcttgtgtcc cccctcacaa tttgccacgt
     1501 gatcgatgcc aaaagcccct tttatgacct atcccagcga agcatgcaaa ctgaacagtt
     1561 cgagattgtc gtcatcctag aaggcattgt ggaaacaact gggatgactt gtcaagctcg
     1621 aacatcatat actgaagatg aagttctttg gggtcatcgt ttttttcctg taatttcctt
     1681 agaagaggga ttctttaaag ttgattactc ccagttccac gcaacatttg aagtccccac
     1741 cccaccttac agtgtgaaag agcaggagga aatgcttctc atgtcgtccc ctttaatagc
     1801 accagccata actaacagca aagaaagaca taattctgtg gaatgcttag atggactaga
     1861 tgatattact acaaaactac catctaagct gcagaaaatt actggaagag aagactttcc
     1921 caaaaaactc ttgaggatga gttctacaac ttcagaaaaa gcctacagct tgggagactt
     1981 gcccatgaaa cttcaacgaa taagttcagt tccgggcaac tcagaagaaa aactggtatc
     2041 taaaaccacc aagatgttat ctgatcccat gagccagtct gtggctgatt tgccaccaaa
     2101 gcttcaaaag atggctggag gagcagctag gatggaaggg aaccttccag ccaaattaag
     2161 aaaaatgaac tctgatcgct tcacactcga ggactacaaa gaccatgacg gtgattataa
     2221 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg atatctcgac
     2281 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2341 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2401 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     2461 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     2521 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     2581 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     2641 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     2701 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     2761 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     2821 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg
     2881 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     2941 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     3001 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     3061 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     3121 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat
     3181 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3241 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3301 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     3361 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3421 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3481 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3541 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3601 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3661 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3721 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3781 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3841 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3901 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3961 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4021 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4081 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4141 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4201 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4261 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4321 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4381 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4441 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4501 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4561 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4621 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4681 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4741 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4801 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     4861 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4921 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4981 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5041 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5101 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     5161 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5221 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5281 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5341 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5401 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5461 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5521 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5581 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5641 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5701 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     5761 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5821 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5881 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5941 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6001 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6061 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6121 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6181 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6241 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6301 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6361 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6421 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6481 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6541 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6601 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6661 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6721 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6781 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6841 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6901 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6961 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7021 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7081 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7141 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7201 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7261 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7321 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
     7381 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     7441 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     7501 ttagg
//
