LOCUS Exported 7699 bp ds-DNA circular SYN 23-JUN-2023 DEFINITION Expresses Gsdmd in mammalian cells. ACCESSION . VERSION . KEYWORDS Flag-Gsdmd SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7699) AUTHORS Liu X, Zhang Z, Ruan J, Pan Y, Magupalli VG, Wu H, Lieberman J TITLE Inflammasome-activated gasdermin D causes pyroptosis by forming membrane pores. JOURNAL Nature. 2016 Jul 6;535(7610):153-8. doi: 10.1038/nature18629. PUBMED 27383986 REFERENCE 2 (bases 1 to 7699) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 23, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7699 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 126..148 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 140..157 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 141..157 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 318..697 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 698..901 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 852..872 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 945..968 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" polyA_signal 2458..3080 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2657..2676) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" primer_bind complement(3104..3124) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3122..3419 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind 3351..3370 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3473..4267 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3527..3546) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4137..4156 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4919..5053 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4956..4975) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5010..5029 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5058..5077) /label=T7 /note="T7 promoter, forward primer" promoter complement(5059..5077) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5091..5107) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5091..5107) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5104..5126) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5115..5131 /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5139..5169) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5184..5205 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5322..5339) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5493..6081) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5573..5592) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6252..7112) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6875..6894 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7113..7217) /gene="bla" /label=AmpR promoter rep_origin 7244..7699 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7331..7350) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7541..7562 /label=F1ori-F /note="F1 origin, forward primer" 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 attaattcag atcgatctac catggactac aaagacgatg 961 acgacaagct tgcggccgca atgccatcgg cctttgagaa agtggtcaag aatgtgatca 1021 aggaggtaag cggcagcaga ggcgatctca ttccggtgga cagcctgcgg aactccacca 1081 gcttcaggcc ctactgcctt ctgaacagga aattttcaag ctcaaggttc tggaaacccc 1141 gttattcatg tgtcaacctg tcaatcaagg acatcctgga gcccagtgct ccagaaccag 1201 aaccggagtg ttttggctcc ttcaaagtct ctgatgtcgt cgatgggaac attcagggca 1261 gagtgatgtt gtcaggcatg ggagaaggga aaatttctgg tggggctgca gtgtctgaca 1321 gttccagtgc ctccatgaat gtgtgtatac tgcgtgtgac tcagaagacc tgggagacca 1381 tgcagcatga aaggcacctt cagcagcctg agaacaaaat cctgcaacag cttcggagtc 1441 gtggggatga cctgtttgtg gtgaccgagg tgctgcagac aaaggaggaa gtgcagatca 1501 ctgaggtcca cagccaagag ggctcaggcc agtttacgct gcctggagct ttatgcttga 1561 agggtgaagg caagggccac caaagccgga agaagatggt gaccattcct gcaggcagca 1621 tcctggcatt ccgagtggcc caactgctta ttggctctaa atgggatatc cttctcgtct 1681 cagatgagaa acagaggacc tttgagccct cctcaggtga cagaaaagca gtgggccaga 1741 ggcaccatgg cctcaatgtg cttgctgcgc tttgttccat cggaaagcag ctcagtctcc 1801 tgtcagatgg gattgatgag gaggaattaa ttgaggcggc agacttccag ggcctgtatg 1861 ctgaggtgaa ggcttgctcc tcagaactgg agagcttgga aatggagttg agacaacaga 1921 tactggtgaa catcggaaag attttacagg accagcccag catggaagcc ttagaggcct 1981 cactagggca gggcctgtgc agtggcggcc aggtggagcc tctggacggc ccagctggct 2041 gcatccttga gtgtctggtg cttgactctg gagaactggt gccggaactc gcagccccta 2101 tcttctacct gctgggagca ctggctgtgc tgagtgaaac ccagcagcag ctgctagcta 2161 aggctctgga gacaacggtg ctgtcaaagc agctggagtt ggtgaagcac gtcttggaac 2221 agagcacccc gtggcaggag cagagttctg tgtccctgcc caccgtgctc cttggggact 2281 gctgggatga aaagaatccc acctgggtct tgctagaaga atgtggccta aggctgcagg 2341 tagaatcccc ccaggtgcac tgggaaccaa cgtctctgat ccccacaagt gcgctctatg 2401 cctccctgtt cctattgtca agtctaggcc agaaaccttg ttagtctaga ggatcccggg 2461 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg 2521 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc 2581 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc 2641 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc 2701 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg 2761 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt 2821 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg 2881 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt 2941 ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc 3001 caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact 3061 cagtagatgc ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta 3121 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 3181 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 3241 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 3301 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 3361 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga 3421 ggaattgatc agcttgggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga 3481 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 3541 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 3601 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga 3661 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 3721 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 3781 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 3841 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 3901 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 3961 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga 4021 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 4081 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 4141 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 4201 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 4261 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca 4321 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 4381 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc 4441 gggctcgatc ccctcgcgag ttggttcagc tgctgcctga ggctggacga cctcgcggag 4501 ttctaccggc agtgcaaatc cgtcggcatc caggaaacca gcagcggcta tccgcgcatc 4561 catgcccccg aactgcagga gtggggaggc acgatggccg ctttggtcga cccggacggg 4621 acgctcctgc gcctgataca gaacgaattg cttgcaggca tctcatgagt gtgtcttccc 4681 gttttccgcc tgaggtcact gcgtggatgg agcgctggcg cctgctgcgc gacggcgagc 4741 tgctcaccac ccactcgcca agctggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 4801 ataggctccg ccgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa 4861 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa 4921 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 4981 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 5041 tcatgtctgg atcaattccc tatagtgagt cgtattaaat tcgtaatcat gtcatagctg 5101 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 5161 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 5221 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 5281 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 5341 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 5401 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 5461 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 5521 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 5581 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 5641 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 5701 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 5761 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 5821 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 5881 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 5941 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 6001 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 6061 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 6121 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 6181 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 6241 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 6301 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 6361 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 6421 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 6481 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 6541 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 6601 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 6661 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 6721 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 6781 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 6841 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 6901 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 6961 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 7021 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 7081 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 7141 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 7201 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc 7261 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 7321 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 7381 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 7441 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 7501 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 7561 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 7621 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa 7681 atattaacgc ttacaattt //