LOCUS       V039340                 8989 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039340
VERSION     V039340
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             949..4425
                     /label="KCNH2(NM_000238)"
                     /note="KCNH2(NM_000238)"
                     /gene="KCNH2"
     CDS             4432..4455
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             4462..4551
                     /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    4589..4813
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4859..5287
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5301..5630
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5697..6491
                     /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    6665..6786
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6835..6851)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6859..6875
                     /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(6883..6913)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6928..6949
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7237..7822)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7993..8853)
                     /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(8854..8958)
                     /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 cggcggccgc gatatcaagc ttgccaccat gccggtgcgg
      961 aggggccacg tcgcgccgca gaacaccttc ctggacacca tcatccgcaa gtttgagggc
     1021 cagagccgta agttcatcat cgccaacgct cgggtggaga actgcgccgt catctactgc
     1081 aacgacggct tctgcgagct gtgcggctac tcgcgggccg aggtgatgca gcgaccctgc
     1141 acctgcgact tcctgcacgg gccgcgcacg cagcgccgcg ctgccgcgca gatcgcgcag
     1201 gcactgctgg gcgccgagga gcgcaaagtg gaaatcgcct tctaccggaa agatgggagc
     1261 tgcttcctat gtctggtgga tgtggtgccc gtgaagaacg aggatggggc tgtcatcatg
     1321 ttcatcctca atttcgaggt ggtgatggag aaggacatgg tggggtcccc ggctcatgac
     1381 accaaccacc ggggcccccc caccagctgg ctggccccag gccgcgccaa gaccttccgc
     1441 ctgaagctgc ccgcgctgct ggcgctgacg gcccgggagt cgtcggtgcg gtcgggcggc
     1501 gcgggcggcg cgggcgcccc gggggccgtg gtggtggacg tggacctgac gcccgcggca
     1561 cccagcagcg agtcgctggc cctggacgaa gtgacagcca tggacaacca cgtggcaggg
     1621 ctcgggcccg cggaggagcg gcgtgcgctg gtgggtcccg gctctccgcc ccgcagcgcg
     1681 cccggccagc tcccatcgcc ccgggcgcac agcctcaacc ccgacgcctc gggctccagc
     1741 tgcagcctgg cccggacgcg ctcccgagaa agctgcgcca gcgtgcgccg cgcctcgtcg
     1801 gccgacgaca tcgaggccat gcgcgccggg gtgctgcccc cgccaccgcg ccacgccagc
     1861 accggggcca tgcacccact gcgcagcggc ttgctcaact ccacctcgga ctccgacctc
     1921 gtgcgctacc gcaccattag caagattccc caaatcaccc tcaactttgt ggacctcaag
     1981 ggcgacccct tcttggcttc gcccaccagt gaccgtgaga tcatagcacc taagataaag
     2041 gagcgaaccc acaatgtcac tgagaaggtc acccaggtcc tgtccctggg cgccgacgtg
     2101 ctgcctgagt acaagctgca ggcaccgcgc atccaccgct ggaccatcct gcattacagc
     2161 cccttcaagg ccgtgtggga ctggctcatc ctgctgctgg tcatctacac ggctgtcttc
     2221 acaccctact cggctgcctt cctgctgaag gagacggaag aaggcccgcc tgctaccgag
     2281 tgtggctacg cctgccagcc gctggctgtg gtggacctca tcgtggacat catgttcatt
     2341 gtggacatcc tcatcaactt ccgcaccacc tacgtcaatg ccaacgagga ggtggtcagc
     2401 caccccggcc gcatcgccgt ccactacttc aagggctggt tcctcatcga catggtggcc
     2461 gccatcccct tcgacctgct catcttcggc tctggctctg aggagctgat cgggctgctg
     2521 aagactgcgc ggctgctgcg gctggtgcgc gtggcgcgga agctggatcg ctactcagag
     2581 tacggcgcgg ccgtgctgtt cttgctcatg tgcacctttg cgctcatcgc gcactggcta
     2641 gcctgcatct ggtacgccat cggcaacatg gagcagccac acatggactc acgcatcggc
     2701 tggctgcaca acctgggcga ccagataggc aaaccctaca acagcagcgg cctgggcggc
     2761 ccctccatca aggacaagta tgtgacggcg ctctacttca ccttcagcag cctcaccagt
     2821 gtgggcttcg gcaacgtctc tcccaacacc aactcagaga agatcttctc catctgcgtc
     2881 atgctcattg gctccctcat gtatgctagc atcttcggca acgtgtcggc catcatccag
     2941 cggctgtact cgggcacagc ccgctaccac acacagatgc tgcgggtgcg ggagttcatc
     3001 cgcttccacc agatccccaa tcccctgcgc cagcgcctcg aggagtactt ccagcacgcc
     3061 tggtcctaca ccaacggcat cgacatgaac gcggtgctga agggcttccc tgagtgcctg
     3121 caggctgaca tctgcctgca cctgaaccgc tcactgctgc agcactgcaa acccttccga
     3181 ggggccacca agggctgcct tcgggccctg gccatgaagt tcaagaccac acatgcaccg
     3241 ccaggggaca cactggtgca tgctggggac ctgctcaccg ccctgtactt catctcccgg
     3301 ggctccatcg agatcctgcg gggcgacgtc gtcgtggcca tcctggggaa gaatgacatc
     3361 tttggggagc ctctgaacct gtatgcaagg cctggcaagt cgaacgggga tgtgcgggcc
     3421 ctcacctact gtgacctaca caagatccat cgggacgacc tgctggaggt gctggacatg
     3481 taccctgagt tctccgacca cttctggtcc agcctggaga tcaccttcaa cctgcgagat
     3541 accaacatga tcccgggctc ccccggcagt acggagttag agggtggctt cagtcggcaa
     3601 cgcaagcgca agttgtcctt ccgcaggcgc acggacaagg acacggagca gccaggggag
     3661 gtgtcggcct tggggccggg ccgggcgggg gcagggccga gtagccgggg ccggccgggg
     3721 gggccgtggg gggagagccc gtccagtggc ccctccagcc ctgagagcag tgaggatgag
     3781 ggcccaggcc gcagctccag ccccctccgc ctggtgccct tctccagccc caggcccccc
     3841 ggagagccgc cgggtgggga gcccctgatg gaggactgcg agaagagcag cgacacttgc
     3901 aaccccctgt caggcgcctt ctcaggagtg tccaacattt tcagcttctg gggggacagt
     3961 cggggccgcc agtaccagga gctccctcga tgccccgccc ccacccccag cctcctcaac
     4021 atccccctct ccagcccggg tcggcggccc cggggcgacg tggagagcag gctggatgcc
     4081 ctccagcgcc agctcaacag gctggagacc cggctgagtg cagacatggc cactgtcctg
     4141 cagctgctac agaggcagat gacgctggtc ccgcccgcct acagtgctgt gaccaccccg
     4201 gggcctggcc ccacttccac atccccgctg ttgcccgtca gccccctccc caccctcacc
     4261 ttggactcgc tttctcaggt ttcccagttc atggcgtgtg aggagctgcc cccgggggcc
     4321 ccagagcttc cccaagaagg ccccacacga cgcctctccc taccgggcca gctgggggcc
     4381 ctcacctccc agcccctgca cagacacggc tcggacccgg gcagtctcga ggattacaag
     4441 gatgacgacg ataagggatc cagcgcctgg tcccaccctc agtttgagaa gggcggaggc
     4501 tctggcggcg gaagcggagg atctgcttgg agccaccccc agttcgaaaa gtgatgaggg
     4561 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     4621 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4681 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4741 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     4801 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     4861 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4921 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4981 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5041 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5101 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5161 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5221 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5281 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5341 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5401 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5461 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5521 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     5581 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     5641 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     5701 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5761 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5821 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     5881 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5941 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     6001 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     6061 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     6121 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     6181 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     6241 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     6301 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6361 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6421 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6481 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     6541 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6601 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6661 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6721 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6781 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6841 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6901 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6961 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7021 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7081 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7141 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7201 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7261 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7321 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7381 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7441 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7501 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7561 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7621 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7681 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7741 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     7801 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7861 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7921 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7981 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     8041 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8101 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8161 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8221 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8281 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8341 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8401 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8461 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8521 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8581 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8641 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8701 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8761 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8821 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8881 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8941 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
