LOCUS V039988 6902 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039988 VERSION V039988 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 908..2395 /label="CASP8(NM_001228)" /note="CASP8(NM_001228)" /gene="CASP8" CDS 2405..2422 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" polyA_signal 2469..2693 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2739..3167 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3181..3510 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3577..4371 /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 4545..4666 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4715..4731) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4739..4755 /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(4763..4793) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4808..4829 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5117..5702) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5873..6733) /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(6734..6838) /gene="bla" /label="AmpR promoter" ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc 901 cgccaccatg gacttcagca gaaatcttta tgatattggg gaacaactgg acagtgaaga 961 tctggcctcc ctcaagttcc tgagcctgga ctacattccg caaaggaagc aagaacccat 1021 caaggatgcc ttgatgttat tccagagact ccaggaaaag agaatgttgg aggaaagcaa 1081 tctgtccttc ctgaaggagc tgctcttccg aattaataga ctggatttgc tgattaccta 1141 cctaaacact agaaaggagg agatggaaag ggaacttcag acaccaggca gggctcaaat 1201 ttctgcctac aggttccact tctgccgcat gagctgggct gaagcaaaca gccagtgcca 1261 gacacagtct gtacctttct ggcggagggt cgatcatcta ttaataaggg tcatgctcta 1321 tcagatttca gaagaagtga gcagatcaga attgaggtct tttaagtttc ttttgcaaga 1381 ggaaatctcc aaatgcaaac tggatgatga catgaacctg ctggatattt tcatagagat 1441 ggagaagagg gtcatcctgg gagaaggaaa gttggacatc ctgaaaagag tctgtgccca 1501 aatcaacaag agcctgctga agataatcaa cgactatgaa gaattcagca aaggggagga 1561 gttgtgtggg gtaatgacaa tctcggactc tccaagagaa caggatagtg aatcacagac 1621 tttggacaaa gtttaccaaa tgaaaagcaa acctcgggga tactgtctga tcatcaacaa 1681 tcacaatttt gcaaaagcac gggagaaagt gcccaaactt cacagcatta gggacaggaa 1741 tggaacacac ttggatgcag gggctttgac cacgaccttt gaagagcttc attttgagat 1801 caagccccac gatgactgca cagtagagca aatctatgag attttgaaaa tctaccaact 1861 catggaccac agtaacatgg actgcttcat ctgctgtatc ctctcccatg gagacaaggg 1921 catcatctat ggcactgatg gacaggaggc ccccatctat gagctgacat ctcagttcac 1981 tggtttgaag tgcccttccc ttgctggaaa acccaaagtg ttttttattc aggcttgtca 2041 gggggataac taccagaaag gtatacctgt tgagactgat tcagaggagc aaccctattt 2101 agaaatggat ttatcatcac ctcaaacgag atatatcccg gatgaggctg actttctgct 2161 ggggatggcc actgtgaata actgtgtttc ctaccgaaac cctgcagagg gaacctggta 2221 catccagtca ctttgccaga gcctgagaga gcgatgtcct cgaggcgatg atattctcac 2281 catcctgact gaagtgaact atgaagtaag caacaaggat gacaagaaaa acatggggaa 2341 acagatgcct cagcctactt tcacactaag aaaaaaactt gtcttccctt ctgatgcggc 2401 cgcacatcac catcatcacc attgactcga gtctagaggg cccgtttaaa cccgctgatc 2461 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc 2521 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc 2581 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg 2641 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga 2701 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt 2761 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 2821 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 2881 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 2941 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 3001 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 3061 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc 3121 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat 3181 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 3241 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga 3301 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 3361 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt 3421 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga 3481 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc 3541 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 3601 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 3661 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 3721 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg 3781 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 3841 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 3901 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 3961 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 4021 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 4081 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat 4141 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 4201 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 4261 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 4321 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 4381 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca 4441 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga 4501 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag 4561 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 4621 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac 4681 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt 4741 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg 4801 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt 4861 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 4921 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 4981 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 5041 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 5101 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 5161 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 5221 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 5281 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 5341 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 5401 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 5461 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 5521 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 5581 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 5641 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 5701 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 5761 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 5821 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 5881 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 5941 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 6001 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 6061 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 6121 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 6181 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 6241 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 6301 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 6361 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 6421 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 6481 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 6541 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 6601 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 6661 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 6721 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 6781 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 6841 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg 6901 gt //