LOCUS V028975 7988 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028975 VERSION V028975 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers primer_bind 141..157 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" enhancer 318..697 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 698..901 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 931..996 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 1009..2736 /label="Il1r1(NM_008362)" /note="Il1r1(NM_008362)" /gene="Il1r1" polyA_signal 2747..3369 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" promoter 3411..3708 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3762..4556 /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 5208..5342 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" promoter complement(5348..5366) /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" primer_bind complement(5380..5396) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5404..5420 /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(5428..5458) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5473..5494 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5782..6370) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6541..7401) /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(7402..7506) /gene="bla" /label="AmpR promoter" rep_origin 7533..7961 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" ORIGIN 1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa 181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta 241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg 361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac 721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc 841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa 961 gatcatgaca tcgattacaa ggatgacgat gacaagcttg cggccgcgat ggagaatatg 1021 aaagtgctac tggggctcat ttgtctcatg gtgcctctgc tgtcgctgga gattgacgta 1081 tgtacagaat atccaaatca gatcgttttg tttttatctg taaatgaaat tgatattcgc 1141 aagtgtcctc ttactccaaa taaaatgcac ggcgacacca taatttggta caagaatgac 1201 agcaagaccc ccatatcagc ggaccgggac tccaggattc atcagcagaa tgaacatctt 1261 tggtttgtac ctgccaaggt ggaggactca ggatattact attgtatagt aagaaactca 1321 acttactgcc tcaaaactaa agtaaccgta actgtgttag agaatgaccc tggcttgtgt 1381 tacagcacac aggccacctt cccacagcgg ctccacattg ccggggatgg aagtcttgtg 1441 tgcccttatg tgagttattt taaagatgaa aataatgagt tacccgaggt ccagtggtat 1501 aagaactgta aacctctgct tcttgacaac gtgagcttct tcggagtaaa agataaactg 1561 ttggtgagga atgtggctga agagcacaga ggggactata tatgccgtat gtcctatacg 1621 ttccggggga agcaatatcc ggtcacacga gtaatacaat ttatcacaat agatgaaaac 1681 aagagggaca gacctgttat cctgagccct cggaatgaga cgatcgaagc tgacccagga 1741 tcaatgatac aactgatctg caacgtcacg ggccagttct cagaccttgt ctactggaag 1801 tggaatggat cagaaattga atggaatgat ccatttctag ctgaagacta tcaatttgtg 1861 gaacatcctt caaccaaaag aaaatacaca ctcattacaa cacttaacat ttcagaagtt 1921 aaaagccagt tttatcgcta tccgtttatc tgtgttgtta agaacacaaa tatttttgag 1981 tcggcgcatg tgcagttaat atacccagtc cctgacttca agaattacct catcgggggc 2041 tttatcatcc tcacggctac aattgtatgc tgtgtgtgca tctataaagt cttcaaggtt 2101 gacatagtgc tttggtacag ggactcctgc tctggttttc ttccttcaaa agcttcagat 2161 ggaaagacat acgatgccta tattctttat cccaagaccc tgggagaggg gtccttctca 2221 gacttagata cttttgtttt taaactgttg cctgaggtct tggagggaca gtttggatac 2281 aagctgttca tttatggaag ggatgactat gttggagaag ataccatcga ggttactaat 2341 gaaaatgtaa agaaaagcag gaggctgatt atcattctag tgagagatat gggaggcttc 2401 agctggctgg gccagtcatc tgaagagcaa atagccatat acaatgctct catccaggaa 2461 ggaattaaaa tcgtcctgct tgagttggag aaaatccaag actatgagaa aatgccagat 2521 tctattcagt tcattaagca gaaacacgga gtcatttgct ggtcaggaga ctttcaagaa 2581 agaccacagt ctgcaaagac caggttctgg aaaaacttaa gataccagat gccagcccaa 2641 cggagatcac cattgtctaa acaccgctta ctaaccctgg atcctgtgcg ggacactaag 2701 gagaaactgc cggcagcaac acacttacca ctcggctagg aattccgggt ggcatccctg 2761 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct 2821 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat 2881 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg 2941 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc 3001 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc 3061 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt 3121 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt 3181 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact 3241 ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg 3301 acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc 3361 tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa 3421 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 3481 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 3541 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag 3601 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 3661 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gaattgatca 3721 gcttgggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 3781 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 3841 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 3901 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc 3961 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 4021 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 4081 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 4141 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 4201 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 4261 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga 4321 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 4381 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 4441 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 4501 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 4561 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga 4621 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg 4681 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccaccccg ggctcgatcc 4741 cctcgcgagt tggttcagct gctgcctgag gctggacgac ctcgcggagt tctaccggca 4801 gtgcaaatcc gtcggcatcc aggaaaccag cagcggctat ccgcgcatcc atgcccccga 4861 actgcaggag tggggaggca cgatggccgc tttggtcgac ccggacggga cgctcctgcg 4921 cctgatacag aacgaattgc ttgcaggcat ctcatgagtg tgtcttcccg ttttccgcct 4981 gaggtcactg cgtggatgga gcgctggcgc ctgctgcgcg acggcgagct gctcaccacc 5041 cactcgccaa gctggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 5101 cgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac 5161 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg 5221 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 5281 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga 5341 tcaattccct atagtgagtc gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg 5401 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 5461 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 5521 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 5581 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 5641 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 5701 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 5761 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 5821 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 5881 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 5941 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 6001 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 6061 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 6121 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 6181 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 6241 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 6301 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 6361 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 6421 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 6481 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 6541 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 6601 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 6661 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 6721 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 6781 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 6841 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 6901 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 6961 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 7021 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 7081 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 7141 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 7201 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 7261 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 7321 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 7381 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 7441 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 7501 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc 7561 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc 7621 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 7681 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 7741 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 7801 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 7861 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg 7921 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct 7981 tacaattt //