LOCUS V047562 6781 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V047562 VERSION V047562 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 964..2271 /label="IRF8(NM_001363907)" /note="IRF8(NM_001363907)" /gene="IRF8" CDS 2278..2343 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2381..2605 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2651..3079 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3093..3422 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3489..4283 /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 4457..4578 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4627..4643) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4651..4667 /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(4675..4705) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4720..4741 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5029..5614) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5785..6645) /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(6646..6750) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc 961 accatggcag gtgtcccgga gtccctgaat ctgatgtgtg accggaatgg tggtcggcgg 1021 cttcgacagt ggctgatcga gcagattgac agtagcatgt atccaggact gatttgggag 1081 aatgaggaga agagcatgtt ccggatccct tggaaacacg ctggcaagca agattataat 1141 caggaagtgg atgcctccat ttttaaggcc tgggcagttt ttaaagggaa gtttaaagaa 1201 ggggacaaag ctgaaccagc cacttggaag acgaggttac gctgtgcttt gaataagagc 1261 ccagattttg aggaagtgac ggaccggtcc caactggaca tttccgagcc atacaaagtt 1321 taccgaattg ttcctgagga agagcaaaaa tgcaaactag gcgtggcaac tgctggctgc 1381 gtgaatgaag ttacagagat ggagtgcggt cgctctgaaa tcgacgagct gatcaaggag 1441 ccttctgtgg acgattacat ggggatgatc aaaaggagcc cttccccgcc ggaggcctgt 1501 cggagtcagc tccttccaga ctggtgggcg cagcagccca gcacaggcgt gccgctggtg 1561 acggggtaca ccacctacga cgcgcaccat tcagcattct cccagatggt gatcagcttc 1621 tactatgggg gcaagctggt gggccaggcc accaccacct gccccgaggg ctgccgcctg 1681 tccctgagcc agcctgggct gcccggcacc aagctgtatg ggcccgaggg cctggagctg 1741 gtgcgcttcc cgccggccga cgccatcccc agcgagcgac agaggcaggt gacgcggaag 1801 ctgttcgggc acctggagcg cggggtgctg ctgcacagca gccggcaggg cgtgttcgtc 1861 aagcggctgt gccagggccg cgtgttctgc agcggcaacg ccgtggtgtg caaaggcagg 1921 cccaacaagc tggagcgtga tgaggtggtc caggtcttcg acaccagcca gttcttccga 1981 gagctgcagc agttctataa cagccagggc cggcttcctg acggcagggt ggtgctgtgc 2041 tttggggaag agtttccgga tatggccccc ttgcgctcca aactcattct cgtgcagatt 2101 gagcagctgt atgtccggca actggcagaa gaggctggga agagctgtgg agccggctct 2161 gtgatgcagg cccccgagga gccgccgcca gaccaggtct tccggatgtt tccagatatt 2221 tgtgcctcac accagagatc atttttcaga gaaaaccaac agatcaccgt cctcgaggac 2281 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac 2341 aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag 2401 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact 2461 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt 2521 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat 2581 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg 2641 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 2701 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 2761 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 2821 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 2881 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 2941 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 3001 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa 3061 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc 3121 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 3181 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3241 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 3301 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 3361 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 3421 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat 3481 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 3541 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 3601 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 3661 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 3721 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 3781 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 3841 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 3901 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 3961 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca 4021 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 4081 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 4141 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 4201 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 4261 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac 4321 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 4381 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 4441 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag 4501 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 4561 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta 4621 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 4681 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 4741 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 4801 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 4861 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 4921 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 4981 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 5041 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 5101 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 5161 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 5221 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 5281 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 5341 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 5401 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 5461 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 5521 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa 5581 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 5641 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 5701 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 5761 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 5821 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 5881 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 5941 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 6001 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 6061 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 6121 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 6181 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 6241 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 6301 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 6361 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 6421 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 6481 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 6541 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 6601 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 6661 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 6721 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 6781 c //