LOCUS       sequence-479199-        7762 bp    DNA     circular SYN 22-JUL-2025
DEFINITION  Expression of N-terminal FLAG tagged human gasdermin C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7762)
  AUTHORS   LaRock DL, Johnson AF, Wilde S, Sands JS, Monteiro MP, LaRock CN
  TITLE     Group A Streptococcus induces GSDMA-dependent pyroptosis in 
            keratinocytes.
  JOURNAL   Nature. 2022 May;605(7910):527-531. doi: 10.1038/s41586-022-04717-x.
            Epub 2022 May 11.
  PUBMED    35545676
REFERENCE   2  (bases 1 to 7762)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41586-022-04717-x"; journalName: "Nature"; date: 
            "2022-05"; volume: "605"; issue: "7910"; pages: "527-531"
COMMENT     Sequence Label: pCMV-Flag-hGSDMC
FEATURES             Location/Qualifiers
     source          1..7762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1005..1024
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1005..1023
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1029..1163
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1053..1072)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1107..1126
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1815..2609)
                     /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(1926..1945)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2536..2555
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2663..2960)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     2958..2978
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    3002..3624
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3406..3425
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             3677..3688
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(5177..5200)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(5244..5447)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5273..5293)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5448..5827
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(5988..6005)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5988..6004)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5997..6019)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(6145..6600)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6282..6303)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6494..6513
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6627..6731
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6732..6800,6801..7592)
                     /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     complement(6950..6969)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga catgattacg aatttaatac gactcactat
     1021 agggaattga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     1081 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1141 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1201 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
     1261 attatgatcg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttccagc
     1321 ttggcgagtg ggtggtgagc agctcgccgt cgcgcagcag gcgccagcgc tccatccacg
     1381 cagtgacctc aggcggaaaa cgggaagaca cactcatgag atgcctgcaa gcaattcgtt
     1441 ctgtatcagg cgcaggagcg tcccgtccgg gtcgaccaaa gcggccatcg tgcctcccca
     1501 ctcctgcagt tcgggggcat ggatgcgcgg atagccgctg ctggtttcct ggatgccgac
     1561 ggatttgcac tgccggtaga actccgcgag gtcgtccagc ctcaggcagc agctgaacca
     1621 actcgcgagg ggatcgagcc cggggtgggc gaagaactcc agcatgagat ccccgcgctg
     1681 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     1741 ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa
     1801 ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa
     1861 tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct
     1921 tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg
     1981 ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca
     2041 tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag cctggcgaac
     2101 agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg
     2161 gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag
     2221 gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg
     2281 gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag
     2341 tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc
     2401 agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga caggtcggtc
     2461 ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag
     2521 ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa
     2581 cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca
     2641 gatcccaagc tgatcaattc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     2701 ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     2761 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     2821 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     2881 tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     2941 gcctggggac tttccacacc ctaactgaca cacattccac agccaagctg gccgcgtacc
     3001 caattcaaca ggcatctact gagtggaccc aacgcatgag aggacagtgc caagcaagca
     3061 actcaaatgt cccaccggtt gggcatggcc aggtagccta tgctgtgtct ggacgtcctc
     3121 ctgctggtat agttatttta aaatcagaag gacagggaag ggagcagtgg ttcacgcctg
     3181 taatcccagc aatttgggag gccaaggtgg gtagatcacc tgagattagg agttggagac
     3241 cagcctggcc aatatggtga aaccccgtct ctaccaaaaa aacaaaaatt agctgagcct
     3301 ggtcatgcat gcctggaatc ccaacaactc gggaggctga ggcaggagaa tcgcttgaac
     3361 ccaggaggcg gagattgcag tgagccaaga ttgtgccact gcactccagc ttggttccca
     3421 atagaccccg caggccctac aggttgtctt cccaacttgc cccttgctcc ataccacccc
     3481 cctccacccc ataatattat agaaggacac ctagtcagac aaaatgatgc aacttaattt
     3541 tattaggaca aggctggtgg gcactggagt ggcaacttcc agggccagga gaggcactgg
     3601 ggaggggtca cagggatgcc acccgggatc ctctagatta ggcctcagcc agctgctgca
     3661 gcaacgagag agtcccatag agggcagaca ggggcatctt tgcttctaca tcccaggttg
     3721 acctggggtt atccagctcc atcctaaggc cacactcctc cagcaggcca taggtgatgg
     3781 ccaaaccctc actctggagt ggggcgagga gctcaggttt gagggtgaag ggaatgctcc
     3841 aggggtatct gaagtttggc tccaggatgc tccttaccag ctcctgttgc tgaagcagga
     3901 tcctcttctc catggaacag gccagcaaat cgtgttggaa gtcactcagc accattatgg
     3961 cttcaaggag ataaagaatg gggtcctttg ggttaaacca tgcatggttt gaatcctgtt
     4021 gaagtttctt taggatggca ccaccagggc catccaaatg acctgagctg tccaattcca
     4081 gcatgttcat caggtcctgt agagcccctc tgtctctgag catggccagg atactgtaga
     4141 acatgacatc ctgaacatcc tttgagagct gagccagtgt ctttattttc tggaaaacct
     4201 cctcttgtag atgcttgaaa ttttgccaga agggttcctc tattcttcct actgggagga
     4261 tgtgaacttg ttcgtatttt ggcaaatgcc cgctcaaaaa ttgctgtgtc agaaatagct
     4321 ctggttttaa cttcatatca ttggatgagg cattgaagag ggttggagag atggtatgaa
     4381 atgatggtag caacccctca ctcctcgcag cacagtagcc taccatttcg gaaatttcgt
     4441 actcatcttg aaaggttctc tgttcatcat catctgagat gagaatggct ttctccttga
     4501 taaccagctg ctttctctta taagccatca ccatgccttt ctgaagagtc agcgccttct
     4561 tcttcactct gagactctct ccttggcctt gacccttgcc ataggtaatc caaagagcaa
     4621 ttttccctaa aatattcaca ctactgctat cgtacagcac agtattgttg atcagttcaa
     4681 cagcctctgt caccacgtac aggttgtccc ctctcctccg gcactccttc agaaatgatg
     4741 gctctggatc caacagtttc cttttttgaa agtcttccag gtttggtgat gggatggtaa
     4801 caatttgaaa ctcgagggag catccatggt ccacagaggc ctcccctgac acactcactt
     4861 ctataccaac attcacaccc atgtcagcct tatgcttctg gatcataatg tcactgaagt
     4921 ggaacggtcc tgtcacaaca gtttctagga ctgaagaact tggctccagg atgtcattga
     4981 gggagaattc aactggaaca tagtcagatt gttcccaaaa tgatgaacga gaatccttct
     5041 tctttcgtaa tataacaaac tgacgtaatt tggtggcact caataggtat ttgacaggtg
     5101 tcaggtcttt gcttccaatc tctttgacca aatttttgct aatgcgttcc aacatggagg
     5161 gcattgcggc cgcaagcttg tcgtcatcgt ctttgtagtc catggtagat cgatctgaat
     5221 taattctgac ggttcactaa acgagctctg cttatataga cctcccaccg tacacgccta
     5281 ccgcccattt gcgtcaacgg ggcggggtta ttacgacatt ttggaaagtc ccgttgattt
     5341 tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag
     5401 tcaaaccgct atccacgccc attggtgtac tgccaaaacc gcatcaccat ggtaatagcg
     5461 atgactaata cgtagatgta ctgccaagta ggaaagtccc gtaaggtcat gtactgggca
     5521 taatgccagg cgggccattt accgtcattg acgtcaatag ggggcggact tggcatatga
     5581 tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa
     5641 tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg
     5701 gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc
     5761 atatatgggc tatgaactaa tgaccccgta attgattact attaataact agtcaataat
     5821 caatgtcaac atggcggtca tattggacat gagccaatat aaatgtacat attatgatat
     5881 agatacaacg tatgcaatgg ccaatagcca atattgattt atgctatata accaatgact
     5941 aatatggcta attgccaata ttgattcaat gtatagatca gcttggcact ggccgtcgtt
     6001 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     6061 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     6121 ttgcgcagcc tgaatggcga atggaaattg taagcgttaa tattttgtta aaattcgcgt
     6181 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     6241 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     6301 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     6361 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     6421 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     6481 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     6541 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6601 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6661 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6721 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     6781 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6841 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6901 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6961 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     7021 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     7081 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     7141 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     7201 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     7261 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     7321 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     7381 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7441 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7501 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7561 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     7621 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     7681 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7741 tagaaaagat caaaggatct tc
//
