LOCUS V019549 6554 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019549 VERSION V019549 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..1234 /label="Rheb(NM_053075)" /note="Rheb(NM_053075)" /gene="Rheb" CDS 1241..1306 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1330..1918 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 1984..2032 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2234..2662 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2676..3005 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3072..3866 /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 4042..4163 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4212..4228) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4236..4252 /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(4260..4290) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4305..4326 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4614..5202) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5373..6233) /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(6234..6338) /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 ccatgcctca gtccaagtcc cggaagatcg ccatcctggg 721 ctaccggtct gtgggaaagt cctcattgac aattcagttt gttgaaggcc aatttgttga 781 ttcctacgat ccaaccatag agaacacgtt caccaagttg atcacggtaa atggtcaaga 841 gtatcatctt cagcttgtag acacagcggg gcaggatgaa tattccattt ttcctcagac 901 atactccata gatattaatg gttatattct tgtgtattct gttacatcaa tcaaaagttt 961 tgaagtaatt aaagttatcc atggcaagtt gttggatatg gtggggaaag tgcagatacc 1021 tattatgttg gttggaaata agaaggacct gcatatggaa agggtgatca gctatgaaga 1081 aggaaaggct ttggcagaat cttggaatgc agcttttttg gaatcttctg ctaaagaaaa 1141 tcaaactgct gttgatgttt ttaaaaggat aattttggaa gcagaaaaga ttgatggagc 1201 agcttcacaa ggaaagtctt cgtgctcggt gatgctcgag gactacaaag accatgacgg 1261 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga 1321 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 1381 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 1441 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 1501 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 1561 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 1621 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 1681 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca 1741 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 1801 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 1861 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 1921 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta 1981 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc 2041 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa 2101 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg 2161 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct 2221 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 2281 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 2341 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta 2401 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 2461 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 2521 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 2581 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt 2641 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 2701 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 2761 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 2821 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 2881 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 2941 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 3001 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 3061 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 3121 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 3181 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 3241 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 3301 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 3361 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 3421 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 3481 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 3541 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 3601 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 3661 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 3721 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 3781 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 3841 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa 3901 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 3961 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 4021 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 4081 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 4141 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 4201 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 4261 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 4321 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 4381 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 4441 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 4501 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 4561 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 4621 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 4681 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 4741 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 4801 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 4861 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 4921 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 4981 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 5041 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 5101 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 5161 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 5221 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 5281 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 5341 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 5401 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 5461 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 5521 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 5581 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 5641 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 5701 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 5761 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 5821 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 5881 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 5941 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 6001 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 6061 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 6121 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 6181 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 6241 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 6301 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 6361 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct 6421 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 6481 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 6541 agaatctgct tagg //