LOCUS V024559 7545 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024559 VERSION V024559 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 705..2201 /label="CLK2(NM_001294338)" /note="CLK2(NM_001294338)" /gene="CLK2" CDS 2208..2297 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 2321..2909 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2975..3023 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3225..3653 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3667..3996 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4063..4857 /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 5033..5154 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5203..5219) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5227..5243 /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(5251..5281) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5296..5317 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5605..6193) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6364..7224) /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(7225..7329) /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 taccgagctc ggatccacta gtccagtgtg gtggaatcgc caccatgccg catcctcgaa 721 ggtaccactc ctcagagcga ggcagccggg ggagttaccg tgaacactat cggagccgaa 781 agcataagcg acgaagaagt cgctcctggt caagtagtag tgaccggaca cgacggcgtc 841 ggcgagagga cagctaccat gtccgttctc gaagcagtta tgatgatcgt tcgtccgacc 901 ggagggtgta tgaccggcga tactgtggca gctacagacg caacgattat agccgggatc 961 ggggagatgc ctactatgac acagactatc ggcattccta tgaatatcag cgggagaaca 1021 gcagttaccg cagccagcgc agcagccgga ggaagcacag acggcggagg aggcgcagcc 1081 ggacatttag ccgctcatct tcgcagcaca gcagccggag agccaagagt gtagaggacg 1141 acgctgaggg ccacctcatc taccacgtcg gggactggct acaagagcga tatgaaatcg 1201 ttagcacctt aggagagggg accttcggcc gagttgtaca atgtgttgac catcgcaggg 1261 gtggggctcg agttgccctg aagatcatta agaatgtgga gaagtacaag gaagcagctc 1321 gacttgagat caacgtgcta gagaaaatca atgagaaaga ccctgacaac aagaacctct 1381 gtgtccagat gtttgactgg tttgactacc atggccacat gtgtatctcc tttgagcttc 1441 tgggccttag caccttcgat ttcctcaaag acaacaacta cctgccctac cccatccacc 1501 aagtgcgcca catggccttc cagctgtgcc aggctgtcaa gttcctccat gataacaagc 1561 tgacacatac agacctcaag cctgaaaata ttctgtttgt gaattcagac tatgagctca 1621 cctacaacct agagaagaag cgagatgagc gcagtgtgaa gagcacagct gtgcgggtgg 1681 tagactttgg cagtgccacc tttgaccatg agcaccatag caccattgtc tccactcgcc 1741 attaccgagc accagaagtc atccttgagt tgggctggtc acagccttgt gatgtgtgga 1801 gtataggctg catcatcttt gaatactatg tgggattcac cctcttccag acccatgaca 1861 acagagagca tctagccatg atggaaagga tcttgggtcc tatcccttcc cggatgatcc 1921 gaaagacaag aaagcagaaa tatttttacc ggggtcgcct ggattgggat gagaacacat 1981 cagctgggcg ctatgttcgt gagaactgca aaccgctgcg gcggtatctg acctcagagg 2041 cagaggaaca ccaccagctc ttcgatctga ttgaaagcat gctagagtat gaaccagcta 2101 agcggctgac cttgggtgaa gcccttcagc atcctttctt cgcccgcctt cgggctgagc 2161 cgcccaacaa gttgtgggac tccagtcggg atatcagtcg gctcgaggaa caaaaactaa 2221 tatcagagga agatttggaa cagaaactaa tcagtgaaga agatttagag caaaagttaa 2281 tttccgagga ggacttatag tgagggcccg atatctcgac aatcaacctc tggattacaa 2341 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata 2401 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc 2461 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg 2521 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac 2581 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat 2641 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt 2701 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat 2761 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc 2821 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag 2881 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg 2941 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg 3001 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt 3061 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc 3121 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc 3181 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg 3241 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 3301 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 3361 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct 3421 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 3481 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 3541 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat 3601 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg 3661 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 3721 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 3781 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 3841 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 3901 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag 3961 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc 4021 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 4081 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 4141 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 4201 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg 4261 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 4321 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 4381 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 4441 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 4501 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 4561 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg 4621 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 4681 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 4741 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 4801 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 4861 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 4921 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 4981 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 5041 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 5101 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 5161 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 5221 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 5281 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 5341 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 5401 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 5461 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 5521 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 5581 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 5641 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 5701 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 5761 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct 5821 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 5881 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 5941 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 6001 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat 6061 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 6121 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 6181 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 6241 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 6301 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 6361 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 6421 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 6481 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 6541 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 6601 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 6661 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 6721 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 6781 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 6841 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 6901 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 6961 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 7021 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 7081 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 7141 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 7201 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 7261 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 7321 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc 7381 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat 7441 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca 7501 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg //