LOCUS V024442 7376 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024442 VERSION V024442 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 683..2023 /label="TBL2(NM_012453)" /note="TBL2(NM_012453)" /gene="TBL2" CDS 2063..2128 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2152..2740 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2806..2854 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3056..3484 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3498..3827 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3894..4688 /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 4864..4985 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5034..5050) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5058..5074 /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(5082..5112) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5127..5148 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5436..6024) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6195..7055) /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(7056..7160) /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 ggatccgcca ccatggagct ctcgcagatg tcggagctca tggggctgtc 721 ggtgttgctt gggctgctgg ccctgatggc gacggcggcg gtagcgcggg ggtggctgcg 781 cgcgggggag gagaggagcg gccggcccgc ctgccaaaaa gcaaatggat ttccacctga 841 caaatcttcg ggatccaaga agcagaaaca atatcagcgg attcggaagg agaagcctca 901 acaacacaac ttcacccacc gcctcctggc tgcagctctg aagagccaca gcgggaacat 961 atcttgcatg gactttagca gcaatggcaa atacctggct acctgtgcag atgatcgcac 1021 catccgcatc tggagcacca aggacttcct gcagcgagag caccgcagca tgagagccaa 1081 cgtggagctg gaccacgcca ccctggtgcg cttcagccct gactgcagag ccttcatcgt 1141 ctggctggcc aacggggaca ccctccgtgt cttcaagatg accaagcggg aggatggggg 1201 ctacaccttc acagccaccc cagaggactt ccctaaaaag cacaaggcgc ctgtcatcga 1261 cattggcatt gctaacacag ggaagtttat catgactgcc tccagtgaca ccactgtcct 1321 catctggagc ctgaagggtc aagtgctgtc taccatcaac accaaccaga tgaacaacac 1381 acacgctgct gtatctccct gtggcagatt tgtagcctcg tgtggcttca ccccagatgt 1441 gaaggtttgg gaagtctgct ttggaaagaa gggggagttc caggaggtgg tgcgagcctt 1501 cgaactaaag ggccactccg cggctgtgca ctcgtttgct ttctccaacg actcacggag 1561 gatggcttct gtctccaagg atggtacatg gaaactgtgg gacacagatg tggaatacaa 1621 gaagaagcag gacccctact tgctgaagac aggccgcttt gaagaggcgg cgggtgccgc 1681 gccgtgccgc ctggccctct cccccaacgc ccaggtcttg gccttggcca gtggcagtag 1741 tattcatctc tacaataccc ggcggggcga gaaggaggag tgctttgagc gggtccatgg 1801 cgagtgtatc gccaacttgt cctttgacat cactggccgc tttctggcct cctgtgggga 1861 ccgggcggtg cggctgtttc acaacactcc tggccaccga gccatggtgg aggagatgca 1921 gggccacctg aagcgggcct ccaacgagag cacccgccag aggctgcagc agcagctgac 1981 ccaggcccaa gagaccctga agagcctggg tgccctgaag aaggaattct gcagatatcc 2041 agcacagtgg cggccgctcg aggactacaa agaccatgac ggtgattata aagatcatga 2101 catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga caatcaacct 2161 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 2221 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 2281 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 2341 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 2401 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 2461 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 2521 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt 2581 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 2641 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 2701 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa 2761 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac 2821 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc 2881 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc 2941 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg 3001 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg 3061 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 3121 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 3181 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct 3241 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 3301 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 3361 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 3421 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 3481 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 3541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 3601 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 3661 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 3721 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 3781 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 3841 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg 3901 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 3961 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 4021 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 4081 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 4141 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 4201 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 4261 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 4321 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 4381 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 4441 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 4501 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 4561 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 4621 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 4681 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca 4741 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc 4801 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac 4861 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 4921 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 4981 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag 5041 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 5101 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 5161 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 5221 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 5281 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 5341 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 5401 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 5461 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 5521 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 5581 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc 5641 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 5701 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 5761 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 5821 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca 5881 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 5941 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 6001 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 6061 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 6121 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 6181 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 6241 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 6301 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 6361 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 6421 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 6481 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 6541 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 6601 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 6661 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 6721 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 6781 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 6841 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 6901 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 6961 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 7021 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 7081 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 7141 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg 7201 ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt 7261 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa 7321 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg //