LOCUS V019259 8372 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019259 VERSION V019259 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 671..3052 /label="UFL1(NM_015323)" /note="UFL1(NM_015323)" /gene="UFL1" CDS 3059..3124 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 3148..3736 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3802..3850 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 4052..4480 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4494..4823 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4890..5684 /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 5860..5981 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6030..6046) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6054..6070 /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(6078..6108) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6123..6144 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6432..7020) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7191..8051) /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(8052..8156) /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 taccgccacc atggcggacg cctgggaaga gattaggcgg ttggcggccg acttccagcg 721 ggcgcagttc gccgaggcca cgcagaggtt gtccgagcgg aactgcattg agattgttaa 781 taaattgatt gctcagaaac agctagaagt agttcataca ctcgatggaa aggaatatat 841 tactccagcc caaattagta aagaaatgag agatgagcta catgtccgag gtggtcgagt 901 aaacattgtt gatctacaac aggtaattaa tgtggacctg attcatattg aaaatagaat 961 tggtgacatt attaaatcag aaaagcatgt tcagttagtg ttgggacaac tgatagatga 1021 gaattatttg gatcggttgg cagaagaggt caatgataaa ttgcaagaaa gtggtcaggt 1081 caccatatca gaactgtgta aaacttatga tcttcctggg aactttctga cacaggcact 1141 aactcagcga cttggtagaa ttatcagtgg acatattgat cttgataata gaggagtaat 1201 ttttacggaa gcttttgtag ctcgacataa agcacgtatc cgtggactat tcagtgctat 1261 tacccggcct acagctgtga attctttgat ttcaaaatat ggatttcagg agcagcttct 1321 ttactctgtg cttgaggaac ttgttaatag cggacgctta cgaggcactg tggttggtgg 1381 gagacaggat aaagctgtgt ttgtccctga catctactcc aggacacaga gtacttgggt 1441 ggattccttt ttcaggcaga atggctatct agaatttgat gctttgtcca gacttggaat 1501 cccagatgct gtaagctaca taaagaaaag atataagact acacaactct tgtttttgaa 1561 agcagcttgt gttggtcaag gacttgtgga tcaagtggaa gcatcagtag aagaagccat 1621 cagctctgga acatgggttg atattgcacc tctgctaccc acttctttat cagttgaaga 1681 tgctgccata ttgcttcagc aggtgatgag ggcattcagc aaacaggcct caactgtagt 1741 ctttagcgac actgttgtag tcagtgaaaa atttataaat gactgtacag aactgttccg 1801 tgagctgatg caccagaaag ctgaaaagga aatgaaaaat aatcctgtgc atttaatcac 1861 tgaagaagat ctgaaacaaa tctccacttt agaaagcgtt agtacaagta aaaaggataa 1921 aaaagatgag cgaagaagga aagcaacaga gggcagtgga agcatgagag gaggaggtgg 1981 gggcaatgcc agagagtaca aaattaaaaa agtcaagaag aaaggaagaa aagatgatga 2041 tagtgatgat gaatctcaat catcccacac tggaaagaag aagccagaga tcagttttat 2101 gttccaggat gagattgaag attttttaag aaaacacata caagatgccc ctgaggagtt 2161 tatttcggaa cttgctgagt acttaataaa acctcttaat aaaacttatc tcgaggtggt 2221 acgttcagta ttcatgtctt caacaacttc tgcttctggg acgggcagaa aacgcacaat 2281 caaggacttg caagaagaag tttcaaacct gtacaataac attaggttat ttgaaaaagg 2341 gatgaagttt tttgcagatg acacacaggc tgctcttacc aaacacttgc tgaagtcagt 2401 gtgtactgat atcactaacc tcattttcaa cttcttagct tcggatttaa tgatggcagt 2461 agacgatcct gcagccatta caagtgaaat aagaaagaaa attttaagta aattatcaga 2521 agaaaccaaa gtagctctta caaaactcca taactctctg aatgaaaaga gcatagaaga 2581 ctttatttct tgtctggatt ctgcagcaga agcttgtgat attatggtga aaaggggaga 2641 caaaaaaagg gaaagacaga tactgttcca acatcgacaa gcactggctg aacagctaaa 2701 ggtcacagaa gaccctgctc ttattctgca cctcacatca gtcctgttgt ttcagttttc 2761 aacccacagc atgctccatg cacctggaag atgtgtccca cagatcattg cttttcttaa 2821 tagtaaaatt ccagaggatc agcatgctct tttggtaaag tatcaaggtt tggttgtaaa 2881 gcagctagtc agtcaaagta agaagactgg gcagggagat tatcccttga ataatgaatt 2941 agacaaagaa caagaagatg ttgccagtac tactcgtaaa gagcttcaag aactttcttc 3001 atccattaaa gaccttgttc tcaaatctag gaaatcatct gtgacggaag agctcgagga 3061 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga 3121 caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga 3181 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg 3241 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc 3301 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc 3361 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt 3421 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt 3481 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg 3541 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg 3601 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg 3661 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt 3721 tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac 3781 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg 3841 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg 3901 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt 3961 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat 4021 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg 4081 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 4141 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 4201 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 4261 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 4321 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 4381 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg 4441 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc 4501 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 4561 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 4621 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 4681 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 4741 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt 4801 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg 4861 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 4921 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 4981 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 5041 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg 5101 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 5161 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 5221 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 5281 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 5341 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 5401 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 5461 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 5521 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 5581 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 5641 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 5701 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc 5761 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 5821 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat 5881 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 5941 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg 6001 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 6061 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 6121 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 6181 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 6241 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 6301 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 6361 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 6421 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 6481 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 6541 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 6601 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag 6661 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 6721 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 6781 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 6841 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg 6901 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 6961 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 7021 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 7081 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 7141 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 7201 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 7261 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 7321 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 7381 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 7441 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 7501 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 7561 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 7621 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 7681 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 7741 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 7801 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 7861 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 7921 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 7981 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 8041 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 8101 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 8161 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt 8221 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 8281 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 8341 tgaccgacaa ttgcatgaag aatctgctta gg //