LOCUS V045357 7102 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045357 VERSION V045357 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 706..1758 /label="EIF2B2(NM_014239)" /note="EIF2B2(NM_014239)" /gene="EIF2B2" CDS 1765..1854 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 1878..2466 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2532..2580 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2782..3210 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3224..3553 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3620..4414 /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 4590..4711 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4760..4776) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4784..4800 /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(4808..4838) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4853..4874 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5162..5750) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5921..6781) /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(6782..6886) /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 ggatccacta gtccagtgtg gtggaattcg ccaccatgcc gggatccgca 721 gcgaagggct cggagttgtc agagaggatc gagagcttcg tggagaccct gaagcggggt 781 ggtgggccgc gcagctccga ggaaatggct cgggagaccc tagggttgct gcgccagatc 841 atcacggacc accgctggag caacgcgggg gagctgatgg agctgatccg cagagagggc 901 aggaggatga cggccgctca gccctccgag accaccgtgg gcaacatggt gcggagagtg 961 ctcaagatta tccgggagga gtatggcaga ctccatggac gcagcgacga gagtgatcag 1021 caggagtccc tgcacaaact gttgacatcc ggaggcctaa acgaggattt cagcttccat 1081 tatgcccaac tccagtccaa catcattgag gcgattaatg agctgctagt ggagctggaa 1141 gggacaatgg agaacattgc agcccaggct ctggagcaca ttcactccaa tgaggtgatc 1201 atgaccattg gcttctcccg aacagtagag gccttcctca aagaggctgc ccgaaagagg 1261 aaattccatg tcattgtagc agagtgtgct cctttctgcc agggtcatga aatggctgtg 1321 aatttgtcca aagcaggtat tgagacaact gtcatgactg atgctgccat ttttgccgtt 1381 atgtcaagag tcaacaaggt gatcattggc acgaagacca tcctggccaa tggggccctg 1441 agagctgtga caggaactca cactctggca ctggcagcaa aacaccattc caccccactc 1501 atcgtctgtg cacctatgtt caaactttct ccacagttcc ccaatgaaga agactcattt 1561 cataagtttg tggctcctga agaagtcctg ccattcacag aaggggacat tctggagaag 1621 gtcagcgtgc attgccctgt gtttgactac gttcccccag agctcattac cctctttatc 1681 tccaacattg gtgggaatgc accttcctac atctaccgcc tgatgagtga actctaccat 1741 cctgatgatc atgttttact cgaggaacaa aaactaatat cagaggaaga tttggaacag 1801 aaactaatca gtgaagaaga tttagagcaa aagttaattt ccgaggagga cttatagtga 1861 gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta 1921 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc 1981 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt 2041 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg 2101 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt 2161 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct 2221 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt 2281 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct 2341 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc 2401 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct 2461 ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac 2521 ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc 2581 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt 2641 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag 2701 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct 2761 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg 2821 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt 2881 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca 2941 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg 3001 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg 3061 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct 3121 cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg 3181 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg 3241 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 3301 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 3361 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc 3421 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc 3481 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct 3541 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac 3601 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc 3661 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc 3721 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc 3781 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg 3841 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt 3901 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc 3961 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga 4021 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga 4081 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct 4141 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc 4201 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt 4261 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg 4321 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat 4381 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga 4441 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg 4501 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 4561 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca 4621 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 4681 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 4741 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 4801 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 4861 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 4921 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 4981 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 5041 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 5101 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 5161 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 5221 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 5281 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 5341 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 5401 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 5461 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 5521 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 5581 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 5641 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 5701 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 5761 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 5821 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 5881 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 5941 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 6001 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 6061 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 6121 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 6181 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 6241 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 6301 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 6361 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 6421 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 6481 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 6541 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 6601 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 6661 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 6721 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 6781 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 6841 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 6901 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag 6961 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga 7021 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa 7081 ttgcatgaag aatctgctta gg //