LOCUS V018845 7406 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018845 VERSION V018845 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 671..2086 /label="CATSPER4(NM_198137)" /note="CATSPER4(NM_198137)" /gene="CATSPER4" CDS 2093..2158 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2182..2770 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2836..2884 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3086..3514 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3528..3857 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3924..4718 /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 4894..5015 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5064..5080) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5088..5104 /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(5112..5142) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5157..5178 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5466..6054) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6225..7085) /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(7086..7190) /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 taccgccacc atgagggata atgaaaaggc ctggtggcag caatggacct cccatacagg 721 cctcgagggg tggggcggga ctcaggagga ccgtatgggg tttggagggg cagtagctgc 781 actgaggggc cgcccctctc ccctgcagag taccattcac gagtcctacg gtcggccaga 841 ggagcaagtg ctcatcaacc gccaggaaat cacgaacaaa gcggacgcct gggacatgca 901 ggagttcatc actcacatgt acatcaagca gctgctccga caccccgcct tccaactgct 961 gctggccctg ctgctggtga tcaatgccat caccatcgct ctccgtacca actcctacct 1021 ggaccagaaa cactatgagt tgttctctac catagatgac attgtgctga ccatccttct 1081 ttgtgaggtt ctccttggct ggctcaatgg cttctggatt ttctggaagg acggctggaa 1141 catcctcaac ttcattatcg tctttatctt gctcttgcgg ttcttcatta atgaaatcaa 1201 tattccctcc atcaactaca ctctcagggc gcttcgtctg gtgcatgtgt gcatggcggt 1261 ggagcccctc gcccggatca tccgcgtcat cctgcagtcg gtgcctgaca tggccaatat 1321 catggtcctc atcctcttct tcatgctggt tttttccgtg tttggagtaa cactctttgg 1381 tgcattcgtg cccaagcatt tccagaacat acaggttgcg ctgtacaccc tcttcatctg 1441 catcacccag gacggctggg tggacatcta cagtgacttc cagacagaga agagggaata 1501 tgcaatggag attgggggtg ccatctactt taccatcttc atcaccatcg gtgccttcat 1561 tggcatcaac ctgttcgtca tcgtggtgac caccaacctg gagcaaatga tgaaggcagg 1621 agagcaggga caacagcaac gaataacctt tagtgagaca ggcgcagagg aagaggagga 1681 gaatgaccag ctgccactgg tgcattgtgt ggtcgcccgc tcggagaaat ctggtctcct 1741 ccaggaaccc cttgcgggag gccccctgtc gaacctctca gaaaacacgt gtgacaactt 1801 ttgcttggtg cttgaggcaa tacaggagaa cctgaggcag tacaaggaga tccgagatga 1861 actcaacatg attgtggagg aggtgcgcgc aatccgcttc aaccaggagc aggagtcaga 1921 ggtgttgaac aggcgctcgt cgacgagcgg gtcgttggag actacgtcat ccaaggacat 1981 ccgccagatg tctcaacagc aagacttgct cagtgcgctc gttagcatgg aaaaggttca 2041 tgactctagc tcacaaatac tccttaaaaa acacaagagc agccacctcg aggactacaa 2101 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta 2161 gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact 2221 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 2281 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 2341 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 2401 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 2461 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 2521 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg 2581 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc 2641 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 2701 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 2761 gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag 2821 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt 2881 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc 2941 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt 3001 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc 3061 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 3121 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 3181 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 3241 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 3301 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 3361 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 3421 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa 3481 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag 3541 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3601 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 3661 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 3721 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 3781 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 3841 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag 3901 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg 3961 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 4021 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 4081 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga 4141 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 4201 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 4261 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 4321 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 4381 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 4441 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 4501 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 4561 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 4621 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 4681 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa 4741 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc 4801 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 4861 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt 4921 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 4981 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct 5041 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 5101 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 5161 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 5221 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 5281 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 5341 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 5401 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 5461 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 5521 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 5581 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 5641 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 5701 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 5761 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 5821 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 5881 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 5941 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 6001 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 6061 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 6121 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 6181 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 6241 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 6301 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 6361 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 6421 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 6481 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 6541 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 6601 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 6661 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 6721 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 6781 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 6841 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 6901 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 6961 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 7021 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 7081 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 7141 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 7201 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc 7261 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt 7321 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg 7381 acaattgcat gaagaatctg cttagg //