LOCUS       sequence-419110-        6720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the bacterial gasdermin (bGSDM) encoded by a 
            Rhodoplanes 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 6720)
  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 6720)
  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_Rhodoplanes-bGSDM
FEATURES             Location/Qualifiers
     source          1..6720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..56)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        124..228
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(229..297,298..1089)
                     /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(447..466)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1260..1848
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1749..1768
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2002..2019
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2034..2173
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2099..2118)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2259..2281
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2275..2466)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4821..4840
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4860..4937)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5143..5162
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5246..5265
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5246..5264
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5265..5289
                     /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             5305..5327
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5347..5364
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6467..6485)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6481..6528
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6703..6708,6709..6720,1..5,6..11))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
       61 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
      121 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      181 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
      241 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
      301 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
      361 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
      421 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
      481 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
      541 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
      601 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
      661 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
      721 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
      781 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
      841 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
      901 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
      961 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1021 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     1081 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1141 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1201 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1261 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1321 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1381 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     1441 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1501 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1561 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1621 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     1681 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     1741 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     1801 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     1861 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     1921 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     1981 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     2041 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     2101 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     2161 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2221 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     2281 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     2341 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     2401 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     2461 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     2521 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     2581 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     2641 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     2701 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     2761 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     2821 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     2881 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga
     2941 cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat
     3001 gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat
     3061 tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc
     3121 cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca
     3181 atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc
     3241 ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga
     3301 tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg
     3361 gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag
     3421 acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     3481 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     3541 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     3601 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     3661 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     3721 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     3781 ctgcccgctt tccagtcggg aaactgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3841 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg
     3901 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
     3961 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
     4021 gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg
     4081 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
     4141 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
     4201 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
     4261 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
     4321 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
     4381 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
     4441 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
     4501 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
     4561 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
     4621 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
     4681 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
     4741 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
     4801 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
     4861 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     4921 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
     4981 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa
     5041 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     5101 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     5161 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     5221 gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat
     5281 aacaattccc ctctagagag ctcttttgtt taactttaag aaggagatat acatatgggt
     5341 tcttctcacc atcaccatca ccatggttca ggagtcaaga ctgagaacaa cgatcatatt
     5401 aatttgaagg tggcggggca ggatggttct gtggtgcagt ttaagattaa gaggcataca
     5461 ccacttagta aactaatgaa agcctattgt gaacgacagg gattgtcaat gaggcagatc
     5521 agattccgat ttgacgggca accaatcaat gaaacagaca cacctgcaca gttggaaatg
     5581 gaggatgaag atacaattga tgtgtttcaa cagcagacgg gtggatccgc agaacagctg
     5641 aatgatggct ttgtgaattt tctgaacagc aaaggttttc agccgattct gctgccgaaa
     5701 ggtggtgttc tgcctccgta tatctatatt ctgggtgcag aaggtagcag cctgtttcag
     5761 tttggtagcc tgaatagcgt tctgccgaaa aaactgggtt ataccaaaaa tgttacctcg
     5821 ccgtttggca aaagcaacat tagcagtaaa ggtggtaatc tgagcgtggg ttttttcaaa
     5881 cagctgctgg aatggtttca tctgggtcaa gcacagctgg atgccggtgg tagcattgaa
     5941 ggtaataccg aatatcgttt tgatggtgtt accacacgtc tggcagaacc gattaaaatc
     6001 ctggcagcac tgaacgatgc aggtattgca ccggcaaaat ttggtgaaaa aattctggcc
     6061 gaaggcaaaa tctatgttgc ctatgaagtt ctgtatgcca ccaccgttaa actgattaaa
     6121 ggtggcgcaa ttaaaggcga ctttaaaatc gaagttagcg ccaatgaaat tgcaagcgca
     6181 aatgcagatg caaaagccga aggtaaaaat gatgaggata gcatctatac agaagatcgt
     6241 ccggttgcat ttgcctttcg tctggcccag ctgctgtatg aagcaaataa acatgaatat
     6301 gccctgaact gggtgaaagt gagcgcagca ggttttagcg aagatgaacc ggaacgtatt
     6361 ctggcagcaa aaggtcaggt tctgaaagtt gcagataaac cgagccgtag ctgataagcg
     6421 gccgctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     6481 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     6541 actatatccg gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta
     6601 cagcatccag ggtgacggtg ccgaggatga cgatgagtgc attgttagat ttcatacacg
     6661 gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga
//
