LOCUS V020328 6464 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020328 VERSION V020328 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..1144 /label="UBE2L5(NM_001355247)" /note="UBE2L5(NM_001355247)" /gene="UBE2L5" CDS 1151..1216 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1240..1828 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 1894..1942 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2144..2572 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2586..2915 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2982..3776 /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 3952..4073 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4122..4138) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4146..4162 /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(4170..4200) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4215..4236 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4524..5112) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5283..6143) /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(6144..6248) /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 ccatggcggc cagcaggagg ctgatgaagg agcttgaaga 721 aatccgcaaa tgtggaatgg aaaacttccg taacatccag gttgatgaag ctaatttatt 781 gacttggcaa gggcttattg ttcctgacaa ccctccgtat aataaggggg ccttcagaat 841 cgaaatcaac tttccagcag agtacccatt caaaccaccg aggatcacat ttaaaacaaa 901 gatctatcac ccgaacatcg acgaaaaggg gcaggtctgt ctgccagtaa ttagtgctga 961 aaactggaag ccagcaacca aaaccgacca agtaatccag tccctcatag cactggtgaa 1021 tgacccgcag cccgagcacc cgcttcgggc tgacctagct gaagaatact ctaacgaccg 1081 taaaaaattc tgtaagaatg ctgaagagtt tacaaagaaa tatggggaaa agcgacctgt 1141 ggacctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa 1201 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa 1261 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 1321 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 1381 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 1441 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 1501 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 1561 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 1621 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 1681 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 1741 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 1801 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga 1861 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc 1921 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc 1981 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc 2041 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct 2101 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 2161 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 2221 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 2281 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc 2341 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 2401 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 2461 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt 2521 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 2581 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 2641 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 2701 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 2761 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 2821 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt 2881 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca 2941 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 3001 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 3061 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 3121 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc 3181 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 3241 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 3301 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 3361 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 3421 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 3481 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga 3541 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 3601 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 3661 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 3721 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 3781 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc 3841 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc 3901 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt 3961 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 4021 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 4081 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 4141 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 4201 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 4261 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 4321 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 4381 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 4441 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 4501 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 4561 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 4621 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 4681 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc 4741 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 4801 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 4861 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 4921 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc 4981 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 5041 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 5101 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 5161 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 5221 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 5281 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 5341 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 5401 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 5461 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 5521 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 5581 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 5641 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 5701 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 5761 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 5821 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 5881 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 5941 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 6001 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 6061 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 6121 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 6181 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 6241 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg 6301 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc 6361 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa 6421 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg //