LOCUS       V027224                 7383 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027224
VERSION     V027224
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..2063
                     /label="KCNJ14(NM_013348)"
                     /note="KCNJ14(NM_013348)"
                     /gene="KCNJ14"
     CDS             2070..2135
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2159..2747
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2813..2861
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3063..3491
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3505..3834
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3901..4695
                     /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    4871..4992
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5041..5057)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5065..5081
                     /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(5089..5119)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5134..5155
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5443..6031)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6202..7062)
                     /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(7063..7167)
                     /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 ccaccatggg cctggccagg gccctacgcc
      781 gcctcagcgg cgccctggat tcgggagaca gccgggcggg cgatgaagag gaggccgggc
      841 ccgggttgtg ccgcaacggg tgggcgccgg caccggtgca gtcacccgtg ggccggcgcc
      901 gcggtcgctt cgtcaagaaa gacgggcact gcaacgtgcg tttcgtaaac ctgggtggcc
      961 agggcgcgcg ctacctgagc gacctgttca ccacatgcgt ggacgtgcgc tggcgctgga
     1021 tgtgcctgct cttctcctgc tccttcctcg cctcctggct gctcttcggc ctggccttct
     1081 ggctcattgc ctcgctgcac ggcgacctgg ccgccccgcc accgcccgcg ccctgcttct
     1141 cacacgtggc cagcttcctg gccgccttcc tcttcgcgct ggagacgcag acgtccatcg
     1201 gctacggcgt gcgcagcgtc accgaggagt gcccggccgc tgtggccgcc gtggtgctgc
     1261 agtgcattgc cggctgcgtg ctcgacgcct tcgtcgtggg tgctgtcatg gccaagatgg
     1321 ccaaacccaa gaagcgcaac gagacgctgg tcttcagcga gaacgccgtc gtggcgctgc
     1381 gcgaccaccg cctctgcctc atgtggcgcg tcggcaacct gcgccgcagc cacctggtcg
     1441 aggcccacgt gcgtgcccag ctgctgcagc cccgtgtgac cccagagggt gagtacatcc
     1501 cgctggacca ccaggatgtg gatgtgggct ttgatggagg caccgatcgt atcttcctcg
     1561 tgtcccccat caccatcgtc catgagatcg actctgccag tcctctgtat gagctaggac
     1621 gtgccgagct ggccagggct gactttgagc tggtggtcat tctcgagggg atggttgagg
     1681 ccacagccat gaccacacag tgtcgctcgt cctacctccc tggtgaactg ctctggggcc
     1741 atcgttttga gccagttctc ttccagcgtg gctcccagta tgaggtcgac tatcgccact
     1801 tccatcgcac ttatgaggtc ccagggacac cggtctgcag tgctaaggag ctggatgaac
     1861 gggcagagca ggcttcccac agcctcaagt ctagtttccc cggctctctg actgcatttt
     1921 gttatgagaa tgaacttgct ctgagctgct gccaggagga agatgaggac gatgagactg
     1981 aggaagggaa tggggtggaa acagaagatg gggctgctag cccccgagtt ctcacaccaa
     2041 ccctggcgct gaccctgcct ccaggatccg actacaaaga ccatgacggt gattataaag
     2101 atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa
     2161 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2221 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2281 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2341 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2401 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2461 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2521 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     2581 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     2641 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     2701 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     2761 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     2821 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     2881 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     2941 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     3001 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     3061 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3121 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3181 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     3241 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3301 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3361 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3421 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3481 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3541 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3601 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3661 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3721 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     3781 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3841 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     3901 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3961 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4021 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4081 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4141 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4201 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4261 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4321 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4381 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4441 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4501 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4561 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4621 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4681 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     4741 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4801 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4861 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4921 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4981 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5041 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5101 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5161 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5221 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5281 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5341 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5401 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5461 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5521 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5581 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     5641 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5701 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5761 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5821 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5881 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5941 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6001 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6061 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6121 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6181 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6241 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6301 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6361 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6421 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6481 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6541 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6601 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6661 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6721 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6781 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6841 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6901 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6961 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7021 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7081 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7141 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     7201 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     7261 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     7321 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     7381 agg
//
