LOCUS       sequence-419098-        6732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the bacterial gasdermin (bGSDM) encoded by a 
            Vitiosangium species. Protein is expressed with an N-terminal SUMO2 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6732)
  AUTHORS   Johnson AG, Wein T, Mayer ML, Duncan-Lowey B, Yirmiya E, 
            Oppenheimer-Shaanan Y, Amitai G, Sorek R, Kranzusch PJ
  TITLE     Bacterial gasdermins reveal an ancient mechanism of cell death.
  JOURNAL   Science. 2022 Jan 14;375(6577):221-225. doi: 
            10.1126/science.abj8432. Epub 2022 Jan 13.
  PUBMED    35025633
REFERENCE   2  (bases 1 to 6732)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.abj8432"; journalName: "Science"; date: 
            "2022-01-14- 14"; volume: "375"; issue: "6577"; pages: "221-225"
COMMENT     Sequence Label: pETSUMO2_Vitiosangium-bGSDM
FEATURES             Location/Qualifiers
     source          1..6732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..29)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        97..201
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(202..270,271..1062)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(420..439)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1233..1821
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1722..1741
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1975..1992
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2007..2146
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2072..2091)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2232..2254
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2248..2439)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4794..4813
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4833..4910)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5116..5135
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5219..5238
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5219..5237
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5238..5262
                     /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)."
     RBS             5278..5300
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5320..5337
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6452..6470)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6466..6513
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6688..6693,6694..6710,6711..6716))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
       61 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
      121 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
      181 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
      241 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
      301 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
      361 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
      421 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
      481 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
      541 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
      601 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
      661 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
      721 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
      781 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
      841 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
      901 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
      961 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     1021 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     1081 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     1141 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     1201 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     1261 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     1321 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     1381 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     1441 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     1501 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     1561 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     1621 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     1681 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     1741 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     1801 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     1861 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     1921 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     1981 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     2041 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     2101 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     2161 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     2221 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     2281 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     2341 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     2401 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     2461 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     2521 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     2581 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     2641 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     2701 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     2761 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     2821 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     2881 caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt
     2941 gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg tttgcgcatt
     3001 cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc cgttagcgag
     3061 gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg
     3121 gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc
     3181 gccgaggcgg cataaatcgc cgtgacgatc agcggtccaa tgatcgaagt taggctggta
     3241 agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc
     3301 atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag
     3361 gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg
     3421 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     3481 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     3541 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     3601 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     3661 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     3721 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaactgt
     3781 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3841 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     3901 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     3961 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     4021 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     4081 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     4141 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     4201 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     4261 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     4321 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     4381 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     4441 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     4501 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     4561 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     4621 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     4681 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     4741 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     4801 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     4861 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     4921 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     4981 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     5041 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     5101 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     5161 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaatta
     5221 atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga gagctctttt
     5281 gtttaacttt aagaaggaga tatacatatg ggttcttctc accatcacca tcaccatggt
     5341 tcaggagtca agactgagaa caacgatcat attaatttga aggtggcggg gcaggatggt
     5401 tctgtggtgc agtttaagat taagaggcat acaccactta gtaaactaat gaaagcctat
     5461 tgtgaacgac agggattgtc aatgaggcag atcagattcc gatttgacgg gcaaccaatc
     5521 aatgaaacag acacacctgc acagttggaa atggaggatg aagatacaat tgatgtgttt
     5581 caacagcaga cgggtggatc cggtctgtgt agcgatccgg caattaccta tctgaaacgt
     5641 ctgggttata acgttgttcg tctgcctcgt gaaggtattc agccgctgca tctgctgggt
     5701 cagcagcgtg gcaccgttga atatctgggt agcctggaaa aactgattac ccagcctccg
     5761 agcgaaccgc ctgcaattac ccgtgatcag gcagcagcag gtattaatgg tcagaaaacc
     5821 gaaaatctga gctttagcat tggcatcaac attctgaaaa gcgttctggc acagtttggt
     5881 gccggtgcag gtattgaagc acagtataat caggcacgta aagtgcgttt tgaatttagc
     5941 aatgttctgg ccgatagcgt tgaaccgctg gcagttggtc agtttctgaa aatggccgaa
     6001 gttgatgcag ataatccggt tctgaaacag tatgtgttag gtaatggtcg cctgtatgtt
     6061 attacccagg tgattaaaag caacgaattt accgttgcag cagaaaaaag cggtggtggt
     6121 agcattcagc tggatgttcc ggaaattcag aaagttgttg gtggtaaact gaaagttgaa
     6181 gcaagcgtta gcagccagag caccgttacc tataaaggtg aaaaacagct ggtgtttggc
     6241 ttcaaatgct ttgaaattgg tgtgaaaaac ggcgaaatta ccctgtttgc aagccagcct
     6301 ggtgcgattg caatggccct ggatgcagcc ggtggtgtta tgccgagcga tagcgcactg
     6361 ctggatgaag gtggtctgct ggatctggaa ggtttttgat aagcggccgc tgctaacaaa
     6421 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     6481 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatat
     6541 cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca tccagggtga
     6601 cggtgccgag gatgacgatg agtgcattgt tagatttcat acacggtgcc tgactgcgtt
     6661 agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc gatgataagc tgtcaaacat
     6721 gagaattctt ga
//
