LOCUS V032908 6968 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032908 VERSION V032908 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..2461 /label="RIPK3(NM_006871)" /note="RIPK3(NM_006871)" /gene="RIPK3" CDS 2471..2488 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" polyA_signal 2535..2759 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2805..3233 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3247..3576 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3643..4437 /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 4611..4732 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4781..4797) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4805..4821 /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(4829..4859) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4874..4895 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5183..5768) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5939..6799) /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(6800..6904) /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 tcgtgcgtca agttatggcc cagcggtgcc cccgccccct tggtgtccat 961 cgaggaactg gagaaccagg agctcgtcgg caaaggcggg ttcggcacag tgttccgggc 1021 gcaacatagg aagtggggct acgatgtggc ggtcaagatc gtaaactcga aggcgatatc 1081 cagggaggtc aaggccatgg caagtctgga taacgaattc gtgctgcgcc tagaaggggt 1141 tatcgagaag gtgaactggg accaagatcc caagccggct ctggtgacta aattcatgga 1201 gaacggctcc ttgtcggggc tgctgcagtc ccagtgccct cggccctggc cgctcctttg 1261 ccgcctgctg aaagaagtgg tgcttgggat gttttacctg cacgaccaga acccggtgct 1321 cctgcaccgg gacctcaagc catccaacgt cctgctggac ccagagctgc acgtcaagct 1381 ggcagatttt ggcctgtcca catttcaggg aggctcacag tcagggacag ggtccgggga 1441 gccagggggc accctgggct acttggcccc agaactgttt gttaacgtaa accggaaggc 1501 ctccacagcc agtgacgtct acagcttcgg gatcctaatg tgggcagtgc ttgctggaag 1561 agaagttgag ttgccaaccg aaccatcact cgtgtacgaa gcagtgtgca acaggcagaa 1621 ccggccttca ttggctgagc tgccccaagc cgggcctgag actcccggct tagaaggact 1681 gaaggagcta atgcagctct gctggagcag tgagcccaag gacagaccct ccttccagga 1741 atgcctacca aaaactgatg aagtcttcca gatggtggag aacaatatga atgctgctgt 1801 ctccacggta aaggatttcc tgtctcagct caggagcagc aataggagat tttctatccc 1861 agagtcaggc caaggaggga cagaaatgga tggctttagg agaaccatag aaaaccagca 1921 ctctcgtaat gatgtcatgg tttctgagtg gctaaacaaa ctgaatctag aggagcctcc 1981 cagctctgtt cctaaaaaat gcccgagcct taccaagagg agcagggcac aagaggagca 2041 ggttccacaa gcctggacag caggcacatc ttcagattcg atggcccaac ctccccagac 2101 tccagagacc tcaactttca gaaaccagat gcccagccct acctcaactg gaacaccaag 2161 tcctggaccc cgagggaatc agggggctga gagacaaggc atgaactggt cctgcaggac 2221 cccggagcca aatccagtaa cagggcgacc gctcgttaac atatacaact gctctggggt 2281 gcaagttgga gacaacaact acttgactat gcaacagaca actgccttgc ccacatgggg 2341 cttggcacct tcgggcaagg ggaggggctt gcagcacccc ccaccagtag gttcgcaaga 2401 aggccctaaa gatcctgaag cctggagcag gccacagggt tggtataatc atagcgggaa 2461 agcggccgca catcaccatc atcaccattg actcgagtct agagggcccg tttaaacccg 2521 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 2581 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 2641 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 2701 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc 2761 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg 2821 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 2881 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 2941 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 3001 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 3061 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 3121 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 3181 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg 3241 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 3301 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 3361 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 3421 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 3481 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag 3541 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc 3601 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 3661 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 3721 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 3781 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg 3841 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 3901 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 3961 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 4021 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 4081 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 4141 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg 4201 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 4261 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 4321 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 4381 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 4441 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 4501 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 4561 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 4621 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 4681 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 4741 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 4801 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 4861 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 4921 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 4981 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 5041 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 5101 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 5161 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 5221 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 5281 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 5341 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 5401 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 5461 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 5521 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 5581 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 5641 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 5701 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 5761 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 5821 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 5881 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 5941 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 6001 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 6061 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 6121 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 6181 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 6241 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 6301 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 6361 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 6421 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 6481 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 6541 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 6601 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 6661 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 6721 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 6781 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 6841 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 6901 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc 6961 cctatggt //