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®, 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 //