LOCUS V021099 8096 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021099 VERSION V021099 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..2743 /label="TGM2(NM_004613)" /note="TGM2(NM_004613)" /gene="TGM2" CDS 2783..2848 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2872..3460 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3526..3574 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3776..4204 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4218..4547 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4614..5408 /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 5584..5705 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5754..5770) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5778..5794 /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(5802..5832) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5847..5868 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6156..6744) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6915..7775) /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(7776..7880) /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 ccatggccga ggagctggtc ttagagaggt gtgatctgga 721 gctggagacc aatggccgag accaccacac ggccgacctg tgccgggaga agctggtggt 781 gcgacggggc cagcccttct ggctgaccct gcactttgag ggccgcaact acgaggccag 841 tgtagacagt ctcaccttca gtgtcgtgac cggcccagcc cctagccagg aggccgggac 901 caaggcccgt tttccactaa gagatgctgt ggaggagggt gactggacag ccaccgtggt 961 ggaccagcaa gactgcaccc tctcgctgca gctcaccacc ccggccaacg cccccatcgg 1021 cctgtatcgc ctcagcctgg aggcctccac tggctaccag ggatccagct ttgtgctggg 1081 ccacttcatt ttgctcttca acgcctggtg cccagcggat gctgtgtacc tggactcgga 1141 agaggagcgg caggagtatg tcctcaccca gcagggcttt atctaccagg gctcggccaa 1201 gttcatcaag aacatacctt ggaattttgg gcagtttgaa gatgggatcc tagacatctg 1261 cctgatcctt ctagatgtca accccaagtt cctgaagaac gccggccgtg actgctcccg 1321 ccgcagcagc cccgtctacg tgggccgggt ggtgagtggc atggtcaact gcaacgatga 1381 ccagggtgtg ctgctgggac gctgggacaa caactacggg gacggcgtca gccccatgtc 1441 ctggatcggc agcgtggaca tcctgcggcg ctggaagaac cacggctgcc agcgcgtcaa 1501 gtatggccag tgctgggtct tcgccgccgt ggcctgcaca gtgctgaggt gcctgggcat 1561 ccctacccgc gtcgtgacca actacaactc ggcccatgac cagaacagca accttctcat 1621 cgagtacttc cgcaatgagt ttggggagat ccagggtgac aagagcgaga tgatctggaa 1681 cttccactgc tgggtggagt cgtggatgac caggccggac ctgcagccgg ggtacgaggg 1741 ctggcaggcc ctggacccaa cgccccagga gaagagcgaa gggacgtact gctgtggccc 1801 agttccagtt cgtgccatca aggagggcga cctgagcacc aagtacgatg cgccctttgt 1861 ctttgcggag gtcaatgccg acgtggtaga ctggatccag caggacgatg ggtctgtgca 1921 caaatccatc aaccgttccc tgatcgttgg gctgaagatc agcactaaga gcgtgggccg 1981 agacgagcgg gaggatatca cccacaccta caaataccca gaggggtcct cagaggagag 2041 ggaggccttc acaagggcga accacctgaa caaactggcc gagaaggagg agacagggat 2101 ggccatgcgg atccgtgtgg gccagagcat gaacatgggc agtgactttg acgtctttgc 2161 ccacatcacc aacaacaccg ctgaggagta cgtctgccgc ctcctgctct gtgcccgcac 2221 cgtcagctac aatgggatct tggggcccga gtgtggcacc aagtacctgc tcaacctcaa 2281 cctggagcct ttctctgaga agagcgttcc tctttgcatc ctctatgaga aataccgtga 2341 ctgccttacg gagtccaacc tcatcaaggt gcgggccctc ctcgtggagc cagttatcaa 2401 cagctacctg ctggctgaga gggacctcta cctggagaat ccagaaatca agatccggat 2461 ccttggggag cccaagcaga aacgcaagct ggtggctgag gtgtccctgc agaacccgct 2521 ccctgtggcc ctggaaggct gcaccttcac tgtggagggg gccggcctga ctgaggagca 2581 gaagacggtg gagatcccag accccgtgga ggcaggggag gaagttaagg tgagaatgga 2641 cctgctgccg ctccacatgg gcctccacaa gctggtggtg aacttcgaga gcgacaagct 2701 gaaggctgtg aagggcttcc ggaatgtcat cattggcccc gccgaattct gcagatatcc 2761 agcacagtgg cggccgctcg aggactacaa agaccatgac ggtgattata aagatcatga 2821 catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga caatcaacct 2881 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 2941 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 3001 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 3061 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 3121 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 3181 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 3241 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt 3301 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 3361 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 3421 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa 3481 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac 3541 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc 3601 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc 3661 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg 3721 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg 3781 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 3841 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 3901 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct 3961 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 4021 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 4081 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 4141 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 4201 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 4261 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 4321 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 4381 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 4441 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 4501 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 4561 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg 4621 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 4681 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 4741 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 4801 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 4861 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 4921 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 4981 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 5041 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 5101 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 5161 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 5221 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 5281 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 5341 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 5401 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca 5461 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc 5521 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac 5581 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 5641 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 5701 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag 5761 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 5821 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 5881 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 5941 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 6001 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 6061 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 6121 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 6181 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 6241 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 6301 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc 6361 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 6421 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 6481 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 6541 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca 6601 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 6661 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 6721 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 6781 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 6841 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 6901 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 6961 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 7021 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 7081 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 7141 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 7201 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 7261 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 7321 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 7381 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 7441 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 7501 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 7561 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 7621 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 7681 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 7741 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 7801 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 7861 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg 7921 ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt 7981 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa 8041 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg //