LOCUS V020339 8402 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020339 VERSION V020339 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..3082 /label="Hnrnpu(NM_016805)" /note="Hnrnpu(NM_016805)" /gene="Hnrnpu" CDS 3089..3154 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 3178..3766 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3832..3880 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 4082..4510 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4524..4853 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4920..5714 /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 5890..6011 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6060..6076) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6084..6100 /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(6108..6138) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6153..6174 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6462..7050) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7221..8081) /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(8082..8186) /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 ccatgagttc ttcgcctgtt aatgtcaaga agctgaaggt 721 gtcggagctg aaggaggagc tcaagaagcg gcgcctctcc gacaagggcc tcaaggccga 781 tctcatggat cgactccagg ccgcgctgga caacgaggca ggaggccgcc ccgccatgga 841 gcccgggaac ggcagtctcg acctaggtgg cgatgcggcc gggcgctcgg gagcgggcct 901 agagcaggag gccgcggctg gcgccgaaga cgacgaggag gaggagggca tcgccgctct 961 ggacggcgac cagatggagc tgggcgagga gaacggggcg gcgggggcgg ctgacgccgg 1021 cgcgatggag gaagaggagg cagcgtcgga agacgagaac ggcgacgacc agggcttcca 1081 ggagggggaa gacgagctcg gcgacgagga ggagggcgcg ggcgacgaga acggtcacgg 1141 ggagcagcag tcccaaccgc cggcagcggc ggcgcagcag cagccttccc agcagcgtgg 1201 tgccggcaag gaggccgcgg gcaagagcag cggccccacc tcgctcttcg cggtgacggt 1261 ggcgccgcca ggggcgaggc agggccaaca gcaggcggga ggagacggca aaacagaaca 1321 gaaaggtgga gataaaaaga gaggcgttaa aagaccgcga gaagatcatg gccgagggta 1381 ttttgagtac atcgaagaaa acaagtacag cagagccaag tctcctcagc cacctgttga 1441 agaagaagac gaacacttcg atgacacagt ggtttgtctt gatacttata attgtgatct 1501 gcattttaaa atctcgagag accgtctgag tgcttcttcc cttactatgg agagttttgc 1561 tttcctgtgg gctggaggaa gagcttccta cggtgtgtca aaaggcaaag tctgctttga 1621 gatgaaggta acagagaaga ttccagtaag acacttatat acaaaagata ttgatataca 1681 tgaagttcgg attggctggt cactaaccac aagtggaatg ttgcttggtg aagaagaatt 1741 ttcttacggg tattctctga aaggaataaa aacatgcaac tgtgagacag aagattatgg 1801 ggagaagttt gatgaaaatg atgtgattac atgctttgct aactttgaaa ctgatgaagt 1861 tgaactctct tatgcgaaga atggacaaga tcttggtgtt gcctttaaga tcagtaagga 1921 agttcttgct gaccggccac tattcccaca tgttctctgc cataactgtg cagttgaatt 1981 taatttcggt caaaaggaaa agccatactt tccaatacct gaagactgta cttttatcca 2041 aaatgtcccc ttagaggacc gagttagagg accaaaagga cctgaagaga agaaggattg 2101 tgaggttgta atgatgattg gcttgccagg agctggaaaa actacctggg ttactaaaca 2161 tgcagctgaa aaccctggga aatacaacat tcttggaaca aatacgatta tggacaagat 2221 gatggtggca ggttttaaga agcaaatggc agatactgga aaactgaaca cactgttgca 2281 gagagcccca cagtgtcttg gcaagtttat tgaaattgct gcccgtaaga agcgaaattt 2341 tattctggat cagacaaatg tgtctgctgc tgcccagaga agaaaaatgt gcctgtttgc 2401 aggcttccag cggaaagctg ttgtagtgtg cccaaaagat gaagactata agcagaggac 2461 acagaagaag gcagaagtag aggggaagga cctaccagaa catgctgtcc tcaagatgaa 2521 aggaaacttc acccttccag aggttgcaga atgctttgat gaaataacct atgttgaact 2581 tcagaaagag gaagcccaaa agcttttgga gcaatataaa gaagaaagca aaaaggcact 2641 gccaccagaa aagaagcaaa acactggctc aaagaaaagc aataagaata agagtggcaa 2701 gaaccagttc aacagaggtg gtggccatag aggccgtgga ggattcaata tgcgaggtgg 2761 caatttcaga ggaggagctc ctgggaatcg tggtggatat aataggagag gcaacatgcc 2821 acagagaggt ggtggcggtg gaagtggtgg aattggctat ccatacccac gtggccctgt 2881 ttttcctggc cgaggtggtt actcaaacag agggaattac aacagaggtg gaatgcccaa 2941 cagagggaac tataaccaga acttcagagg acgaggaaat aatcgtggct acaaaaatca 3001 atctcagggc tacaatcagt ggcagcaggg tcaattctgg ggtcagaagc catggagtca 3061 gcattatcac caaggatatt atctcgagga ctacaaagac catgacggtg attataaaga 3121 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata tctcgacaat 3181 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct 3241 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg 3301 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg 3361 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt 3421 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt 3481 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg 3541 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc 3601 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat 3661 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc 3721 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga 3781 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa 3841 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc 3901 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt 3961 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac 4021 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc 4081 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 4141 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 4201 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt 4261 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 4321 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 4381 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 4441 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 4501 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc 4561 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt 4621 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 4681 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc 4741 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg 4801 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc 4861 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca 4921 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg 4981 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag 5041 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc 5101 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc 5161 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg 5221 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc 5281 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca 5341 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag 5401 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg 5461 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg 5521 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca 5581 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc 5641 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg 5701 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac 5761 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc 5821 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc 5881 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 5941 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 6001 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg 6061 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 6121 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 6181 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 6241 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 6301 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 6361 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 6421 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 6481 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 6541 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 6601 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc 6661 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 6721 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 6781 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 6841 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 6901 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 6961 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 7021 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 7081 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 7141 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 7201 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 7261 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 7321 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 7381 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 7441 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 7501 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 7561 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 7621 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 7681 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 7741 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 7801 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 7861 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 7921 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 7981 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 8041 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 8101 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 8161 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac 8221 ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg 8281 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga 8341 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta 8401 gg //