LOCUS       V045183                 6826 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045183
VERSION     V045183
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             928..2262
                     /label="KCNJ4(NM_004981)"
                     /note="KCNJ4(NM_004981)"
                     /gene="KCNJ4"
     CDS             2269..2292
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2299..2388
                     /codon_start=1
                     /product="two Strep-Tag®II moieties connected by a linker
                     for enhanced binding to Strep-Tactin®,&nbsp;an engineered
                     form of streptavidin (Schmidt et al., 2013)"
                     /transl_table=1
                     /translation="SA,WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK"
                     /label="Twin-Strep-tag"
     polyA_signal    2426..2650
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2696..3124
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3138..3467
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3534..4328
                     /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    4502..4623
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4672..4688)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4696..4712
                     /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(4720..4750)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4765..4786
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5074..5659)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5830..6690)
                     /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(6691..6795)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagctgaatt cgccaccatg cacggacaca gccgcaacgg ccaggcccac
      961 gtgccccggc ggaagcgccg caaccgcttc gtcaagaaga acggccaatg caacgtgtac
     1021 ttcgccaacc tgagcaacaa gtcgcagcgc tacatggcgg acatcttcac cacctgcgtg
     1081 gacacgcgct ggcgctacat gctcatgatc ttctccgcgg ccttccttgt ctcctggctc
     1141 tttttcggcc tcctcttctg gtgtatcgcc ttcttccacg gtgacctgga ggccagccca
     1201 ggggtgcctg cggcgggggg cccggcggcg ggtggtggcg gagcagcccc ggtggccccc
     1261 aagccctgca tcatgcacgt gaacggcttc ctgggtgcct tcctgttctc ggtggagacg
     1321 cagacgacca tcggctatgg gttccggtgc gtgacagagg agtgcccgct ggcagtcatc
     1381 gctgtggtgg tccagtccat cgtgggctgc gtcatcgact ccttcatgat tggcaccatc
     1441 atggccaaga tggcgcggcc caagaagcgg gcgcagacgt tgctgttcag ccaccacgcg
     1501 gtcatttcgg tgcgcgacgg caagctctgc ctcatgtggc gcgtgggcaa cctgcgcaag
     1561 agccacattg tggaggccca cgtgcgggcc cagctcatca agccctacat gacccaggag
     1621 ggcgagtacc tgcccctgga ccagcgggac ctcaacgtgg gctatgacat cggcctggac
     1681 cgcatcttcc tggtgtcgcc catcatcatt gtccacgaga tcgacgagga cagcccgctt
     1741 tatggcatgg gcaaggagga gctggagtcg gaggactttg agatcgtggt catcctggag
     1801 ggcatggtgg aggccacggc catgaccacc caggcccgca gctcctacct ggccagcgag
     1861 atcctgtggg gccaccgctt tgagcctgtg gtcttcgagg agaagagcca ctacaaggtg
     1921 gactactcac gttttcacaa gacctacgag gtggccggca cgccctgctg ctcggcccgg
     1981 gagctgcagg agagtaagat caccgtgctg cccgccccac cgccccctcc cagtgccttc
     2041 tgctacgaga acgagctggc ccttatgagc caggaggaag aggagatgga ggaggaggca
     2101 gctgcggcgg ccgcggtggc cgcaggcctg ggcctggagg cgggttccaa ggaggaggcg
     2161 ggcatcatcc ggatgctgga gttcggcagc cacctggacc tggagcgcat gcaggcttcc
     2221 ctcccgctgg acaacatctc ctaccgcagg gagtctgcca tcctcgagga ttacaaggat
     2281 gacgacgata agggatccag cgcctggtcc caccctcagt ttgagaaggg cggaggctct
     2341 ggcggcggaa gcggaggatc tgcttggagc cacccccagt tcgaaaagtg atgagggccc
     2401 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2461 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2521 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2581 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2641 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     2701 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2761 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2821 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2881 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2941 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3001 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3061 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3121 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3181 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3241 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3301 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3361 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3421 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3481 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3541 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3601 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3661 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3721 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3781 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3841 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3901 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3961 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4021 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4081 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4141 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4201 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4261 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4321 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4381 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4441 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4501 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4561 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4621 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4681 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4741 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4801 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4861 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4921 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4981 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5041 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5101 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5161 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5221 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5281 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5341 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5401 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5461 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5521 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5581 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     5641 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5701 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5761 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5821 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5881 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5941 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6001 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6061 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6121 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6181 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6241 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6301 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6361 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6421 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6481 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6541 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6601 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6661 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6721 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6781 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
