LOCUS V031795 6976 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031795 VERSION V031795 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 706..1632 /label="PPP2CA(NM_002715)" /note="PPP2CA(NM_002715)" /gene="PPP2CA" CDS 1639..1728 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 1752..2340 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2406..2454 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2656..3084 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3098..3427 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3494..4288 /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 4464..4585 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4634..4650) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4658..4674 /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(4682..4712) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4727..4748 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5036..5624) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5795..6655) /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(6656..6760) /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 gtggaattcg ccaccatgga cgagaaggtg 721 ttcaccaagg agctggacca gtggatcgag cagctgaacg agtgcaagca gctgtccgag 781 tcccaggtca agagcctctg cgagaaggct aaagaaatcc tgacaaaaga atccaacgtg 841 caagaggttc gatgtccagt tactgtctgt ggagatgtgc atgggcaatt tcatgatctc 901 atggaactgt ttagaattgg tggcaaatca ccagatacaa attacttgtt tatgggagat 961 tatgttgaca gaggatatta ttcagttgaa acagttacac tgcttgtagc tcttaaggtt 1021 cgttaccgtg aacgcatcac cattcttcga gggaatcatg agagcagaca gatcacacaa 1081 gtttatggtt tctatgatga atgtttaaga aaatatggaa atgcaaatgt ttggaaatat 1141 tttacagatc tttttgacta tcttcctctc actgccttgg tggatgggca gatcttctgt 1201 ctacatggtg gtctctcgcc atctatagat acactggatc atatcagagc acttgatcgc 1261 ctacaagaag ttccccatga gggtccaatg tgtgacttgc tgtggtcaga tccagatgac 1321 cgtggtggtt ggggtatatc tcctcgagga gctggttaca cctttgggca agatatttct 1381 gagacattta atcatgccaa tggcctcacg ttggtgtcta gagctcacca gctagtgatg 1441 gagggatata actggtgcca tgaccggaat gtagtaacga ttttcagtgc tccaaactat 1501 tgttatcgtt gtggtaacca agctgcaatc atggaacttg acgatactct aaaatactct 1561 ttcttgcagt ttgacccagc acctcgtaga ggcgagccac atgttactcg tcgtacccca 1621 gactacttcc tgctcgagga acaaaaacta atatcagagg aagatttgga acagaaacta 1681 atcagtgaag aagatttaga gcaaaagtta atttccgagg aggacttata gtgagggccc 1741 gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta 1801 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta 1861 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt 1921 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg 1981 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt 2041 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag 2101 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc 2161 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc 2221 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc 2281 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc 2341 ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc 2401 tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac 2461 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc 2521 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag 2581 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct 2641 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta 2701 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc 2761 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt 2821 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg 2881 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca 2941 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct 3001 attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga 3061 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa 3121 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 3181 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 3241 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag 3301 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 3361 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt 3421 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg 3481 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt 3541 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt 3601 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc 3661 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc 3721 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga 3781 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat 3841 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca 3901 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga 3961 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc 4021 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat 4081 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga 4141 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg 4201 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt 4261 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc 4321 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt 4381 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 4441 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa 4501 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 4561 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct 4621 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 4681 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 4741 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 4801 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 4861 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 4921 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 4981 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 5041 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 5101 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 5161 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 5221 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 5281 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 5341 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 5401 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 5461 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 5521 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 5581 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 5641 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 5701 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 5761 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 5821 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 5881 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 5941 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 6001 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 6061 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 6121 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 6181 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 6241 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 6301 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 6361 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 6421 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 6481 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 6541 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 6601 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 6661 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 6721 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 6781 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat 6841 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc 6901 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat 6961 gaagaatctg cttagg //