LOCUS V040250 8286 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040250 VERSION V040250 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" promoter 678..696 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 756..2966 /label="DDX50(NM_024045)" /note="DDX50(NM_024045)" /gene="DDX50" CDS 2973..3038 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 3062..3650 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3716..3764 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3966..4394 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4408..4737 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4804..5598 /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 5774..5895 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5944..5960) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5968..5984 /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(5992..6022) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6037..6058 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6346..6934) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7105..7965) /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(7966..8070) /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 gctctctggc taactagaga acccactgct 661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta 721 aacttaagct tggtaccgag ctcgaattcg ccaccatgcc tgggaaactc ctctgggggg 781 acattatgga gctggaagca cccttggagg agtccgagag ccagaagaag gagaggcaaa 841 agagtgacag aaggaagtca aggcaccatt atgactcgga tgagaaatca gaaacaagag 901 aaaatggtgt tacagatgac ctggatgctc ccaaggccaa aaaatctaaa atgaaagaga 961 agctaaatgg agacactgaa gaaggattta atagactttc agatgaattc tccaaatctc 1021 ataagtcaag aagaaaagat ctaccaaatg gagatataga tgaatatgaa aaaaaatcaa 1081 agcgagtatc atctttagat acttctactc ataaatcaag tgataataaa ctagaggaga 1141 ccttaacacg tgaacagaaa gaaggagcct tctccaattt tcctatttct gaagagacta 1201 taaagcttct gaaaggtcga ggggtaacat atctctttcc tattcaagtt aagacctttg 1261 gtcctgtata tgaaggaaaa gatttaatag ctcaagcacg gacaggaaca ggaaagacat 1321 tctcttttgc catcccctta attgaaagac tccaaagaaa tcaagaaaca attaaaaaaa 1381 gccgctcacc aaaggtactt gttttggctc caacaaggga actggcaaac caagtagcca 1441 aagacttcaa agatataact aggaaactca gcgtggcgtg tttttatggt ggaacatcat 1501 atcaaagcca aattaatcat attcgaaatg gtattgacat cttggttgga acacctggtc 1561 gtatcaaaga ccatctgcag agtggccgat tggatctttc taaactgcga catgttgtgc 1621 ttgatgaagt ggatcagatg ttagatttag gtttcgctga acaagttgaa gatattattc 1681 atgaatccta caaaactgat tctgaagaca atcctcagac tttacttttt tctgcaactt 1741 gcccacagtg ggtatacaaa gttgcaaaaa aatacatgaa atccagatat gaacaggttg 1801 accttgttgg aaaaatgact caaaaggctg caactactgt ggaacatttg gccatccagt 1861 gtcattggtc tcagaggcca gcagttattg gagatgtcct tcaagtctac agtgggtctg 1921 aagggagggc tattattttc tgtgagacca agaagaatgt aactgaaatg gccatgaatc 1981 cacacataaa acagaatgcc cagtgtttac atggggacat tgcacagtca caaagagaaa 2041 ttacactaaa aggcttcaga gaaggtagtt ttaaagtttt ggtggcaacc aatgtggctg 2101 cccgtggttt ggacattcct gaagttgacc tggtgattca aagttctcct cctcaggatg 2161 ttgagtccta tatccatcgc tctggacgca caggtagagc tggacggaca gggatttgta 2221 tatgttttta tcaaccaaga gaaagaggtc aactaagata tgtggaacaa aaagcaggaa 2281 ttacttttaa acgtgtaggt gttccttcta caatggattt agttaaatct aaaagcatgg 2341 atgccatcag gtctctggct tccgtttctt acgctgctgt tgattttttc cgaccatcag 2401 ctcagagact gatagaagag aaaggtgcag tggatgcatt ggctgcagct ttagcccaca 2461 tttctggtgc atcaagcttt gaaccacgat ctttgatcac ctctgataag gggtttgtga 2521 ccatgactct ggaaagccta gaggaaatac aggatgtcag ctgtgcttgg aaagaactta 2581 acagaaagct gagtagtaat gcagtgtctc agattaccag aatgtgcctc ctgaaaggaa 2641 atatgggtgt ttgctttgat gttcctacaa ctgagtcaga aaggttacag gcagagtggc 2701 atgattccga ctggatactc tcagtgccag ccaaattacc tgaaattgaa gaatattatg 2761 atggaaacac atcttctaat tccagacaga ggagtggctg gtcaagtggt cgatcaggcc 2821 ggtcaggccg gtcaggtggt cgatctggcg gccggtcagg tagacagagt cgacaaggaa 2881 gtcgctcagg aagtcgacaa gatggtagaa gacgaagtgg gaatagaaat cgatcaagaa 2941 gtgggggcca caaacggagt tttgacggat ccgactacaa agaccatgac ggtgattata 3001 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga 3061 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 3121 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 3181 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 3241 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 3301 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 3361 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 3421 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct 3481 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 3541 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 3601 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg 3661 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca 3721 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg 3781 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg 3841 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc 3901 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta 3961 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 4021 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 4081 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg 4141 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 4201 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 4261 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 4321 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 4381 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 4441 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 4501 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 4561 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 4621 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 4681 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 4741 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 4801 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 4861 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 4921 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 4981 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 5041 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 5101 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 5161 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 5221 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 5281 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 5341 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 5401 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 5461 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 5521 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 5581 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc 5641 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg 5701 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt 5761 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 5821 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 5881 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc 5941 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 6001 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 6061 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 6121 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 6181 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 6241 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 6301 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 6361 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 6421 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 6481 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 6541 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 6601 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 6661 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 6721 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 6781 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 6841 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 6901 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 6961 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 7021 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 7081 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 7141 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 7201 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 7261 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 7321 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 7381 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 7441 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 7501 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 7561 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 7621 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 7681 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 7741 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 7801 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 7861 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 7921 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 7981 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 8041 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 8101 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat 8161 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc 8221 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg 8281 cttagg //