LOCUS V020648 7172 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020648 VERSION V020648 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..1852 /label="Ilf2(NM_026374)" /note="Ilf2(NM_026374)" /gene="Ilf2" CDS 1859..1924 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1948..2536 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2602..2650 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2852..3280 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3294..3623 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3690..4484 /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 4660..4781 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4830..4846) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4854..4870 /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(4878..4908) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4923..4944 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5232..5820) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5991..6851) /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(6852..6956) /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 ccatgagggg tgacagagga cgtggtcgtg gtgggcgctt 721 tggttccaga ggagggcctg gtggagggtt caggcctttt gtgccccata tcccatttga 781 tttttatttg tgtgaaatgg cttttccccg ggtcaagcca gcacctgatg agacctcctt 841 cagtgaagcc ttgctgaaga ggaatcaaga ccttgcccct aattctgctg agcaggcttc 901 catcctttct ttggtgacaa aaatcaacaa tgtgattgac aacttgattg tggccccagg 961 aacatttgaa gtgcaaattg aagaagtccg gcaggtagga tcatataaaa agggaacaat 1021 gactacagga cacaatgtgg ctgacctggt ggtaatacta aagatcctgc caacattgga 1081 agctgttgct gccctgggga acaaagtcgt ggaaagccta agggcacagg atccctcaga 1141 agttttgacc atgctgacta atgaaactgg ctttgaaatt agctcttctg atgctacggt 1201 gaaaattctc attacaacag tgccccccaa tctccgaaaa ctggatccag aactccattt 1261 ggacatcaag gtgttgcaga gcgccttagc agccatccgg catgcccgct ggtttgaaga 1321 gaacgcttct cagtccacag ttaaagttct cataagattg ctaaaggact tacggattcg 1381 ttttcctggc tttgagcctc tcacaccctg gatccttgac ctacttgggc actatgctgt 1441 aatgaacaac cccaccagac agcctttggc cctgaatgtg gcttacaggc gctgcttgca 1501 gattctggct gcaggactgt tcttgcctgg gtcagtgggc atcactgacc cgtgtgagag 1561 tggcaacttc agggtacaca cagttatgac cctagaacaa caggatatgg tctgctacac 1621 agctcagact ctggttcgaa ttctctccca tggaggcttt aggaagatcc ttggccagga 1681 gggggatgcc agctatcttg cttctgaaat atctacctgg gatggagtga ttgtgacacc 1741 ctcagaaaag gcttatgaga agccacctga gaagaaggag ggggaggaag aggaggagaa 1801 cacagaagaa ccacctcaag gagaagaaga agaaagcatg gagactcagg agctcgagga 1861 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga 1921 caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga 1981 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg 2041 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc 2101 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc 2161 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt 2221 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt 2281 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg 2341 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg 2401 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg 2461 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt 2521 tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac 2581 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg 2641 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg 2701 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt 2761 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat 2821 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg 2881 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 2941 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 3001 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 3061 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 3121 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 3181 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg 3241 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc 3301 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 3361 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 3421 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 3481 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 3541 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt 3601 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg 3661 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 3721 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 3781 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 3841 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg 3901 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 3961 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 4021 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 4081 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 4141 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 4201 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 4261 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 4321 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 4381 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 4441 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 4501 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc 4561 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 4621 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat 4681 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 4741 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg 4801 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 4861 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 4921 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 4981 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 5041 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 5101 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 5161 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 5221 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 5281 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 5341 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 5401 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag 5461 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 5521 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 5581 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 5641 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg 5701 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 5761 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 5821 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 5881 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 5941 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 6001 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 6061 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 6121 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 6181 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 6241 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 6301 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 6361 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 6421 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 6481 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 6541 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 6601 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 6661 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 6721 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 6781 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 6841 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 6901 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 6961 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt 7021 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 7081 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 7141 tgaccgacaa ttgcatgaag aatctgctta gg //