LOCUS       sequence-332599-        7390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Purification of catalytically-dead DPP9S from insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7390)
  AUTHORS   Hollingsworth LR, Sharif H, Griswold AR, Fontana P, Mintseris J, 
            Dagbay KB, Paulo JA, Gygi SP, Bachovchin DA, Wu H
  TITLE     DPP9 sequesters the C terminus of NLRP1 to repress inflammasome 
            activation.
  JOURNAL   Nature. 2021 Mar 17. pii: 10.1038/s41586-021-03350-4. doi: 
            10.1038/s41586-021-03350-4.
  PUBMED    33731932
REFERENCE   2  (bases 1 to 7390)
  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"; journalName: "Nature. 
            2021 Mar 17. pii: 10.1038/s41586-021-03350-4. doi: 
            10.1038/s41586-021-03350-4."
COMMENT     Sequence Label: pFastBac HTB DPP9S S730A
FEATURES             Location/Qualifiers
     source          1..7390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..92
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     17..34
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             159..176
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             198..218
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2985..3119
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3022..3041)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3076..3095
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3148..3313
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3497..3951
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3584..3603)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3793..3814
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3977..4081
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4082..4150,4151..4942)
                     /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(4300..4319)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5113..5701
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5602..5621
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5866..5883
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6017..6241
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6308..6841)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7011..7016,7017..7033,7034..7039))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc
       61 gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca
      121 tcgggcgcgg atctcggtcc gaaaccatgt cgtactacca tcaccatcac catcacgatt
      181 acgatatccc aacgaccgaa aacctgtatt ttcagggcgc catgggatcc atggccacca
      241 ccgggacccc aacggccgac cgaggcgacg cagccgccac agatgacccg gccgcccgct
      301 tccaggtgca gaagcactcg tgggacgggc tccggagcat catccacggc agccgcaagt
      361 actcgggcct cattgtcaac aaggcgcccc acgacttcca gtttgtgcag aagacggatg
      421 agtctgggcc ccactcccac cgcctctact acctgggaat gccatatggc agccgagaga
      481 actccctcct ctactctgag attcccaaga aggtccggaa agaggctctg ctgctcctgt
      541 cctggaagca gatgctggat catttccagg ccacgcccca ccatggggtc tactctcggg
      601 aggaggagct gctgagggag cggaaacgcc tgggggtctt cggcatcacc tcctacgact
      661 tccacagcga gagtggcctc ttcctcttcc aggccagcaa cagcctcttc cactgccgcg
      721 acggcggcaa gaacggcttc atggtgtccc ctatgaaacc gctggaaatc aagacccagt
      781 gctcagggcc ccggatggac cccaaaatct gccctgccga ccctgccttc ttctccttca
      841 tcaataacag cgacctgtgg gtggccaaca tcgagacagg cgaggagcgg cggctgacct
      901 tctgccacca aggtttatcc aatgtcctgg atgaccccaa gtctgcgggt gtggccacct
      961 tcgtcataca ggaagagttc gaccgcttca ctgggtactg gtggtgcccc acagcctcct
     1021 gggaaggttc agagggcctc aagacgctgc gaatcctgta tgaggaagtc gatgagtccg
     1081 aggtggaggt cattcacgtc ccctctcctg cgctagaaga aaggaagacg gactcgtatc
     1141 ggtaccccag gacaggcagc aagaatccca agattgcctt gaaactggct gagttccaga
     1201 ctgacagcca gggcaagatc gtctcgaccc aggagaagga gctggtgcag cccttcagct
     1261 cgctgttccc gaaggtggag tacatcgcca gggccgggtg gacccgggat ggcaaatacg
     1321 cctgggccat gttcctggac cggccccagc agtggctcca gctcgtcctc ctccccccgg
     1381 ccctgttcat cccgagcaca gagaatgagg agcagcggct agcctctgcc agagctgtcc
     1441 ccaggaatgt ccagccgtat gtggtgtacg aggaggtcac caacgtctgg atcaatgttc
     1501 atgacatctt ctatcccttc ccccaatcag agggagagga cgagctctgc tttctccgcg
     1561 ccaatgaatg caagaccggc ttctgccatt tgtacaaagt caccgccgtt ttaaaatccc
     1621 agggctacga ttggagtgag cccttcagcc ccggggaaga tgaatttaag tgccccatta
     1681 aggaagagat tgctctgacc agcggtgaat gggaggtttt ggcgaggcac ggctccaaga
     1741 tctgggtcaa tgaggagacc aagctggtgt acttccaggg caccaaggac acgccgctgg
     1801 agcaccacct ctacgtggtc agctatgagg cggccggcga gatcgtacgc ctcaccacgc
     1861 ccggcttctc ccatagctgc tccatgagcc agaacttcga catgttcgtc agccactaca
     1921 gcagcgtgag cacgccgccc tgcgtgcacg tctacaagct gagcggcccc gacgacgacc
     1981 ccctgcacaa gcagccccgc ttctgggcta gcatgatgga ggcagccagc tgccccccgg
     2041 attatgttcc tccagagatc ttccatttcc acacgcgctc ggatgtgcgg ctctacggca
     2101 tgatctacaa gccccacgcc ttgcagccag ggaagaagca ccccaccgtc ctctttgtat
     2161 atggaggccc ccaggtgcag ctggtgaata actccttcaa aggcatcaag tacttgcggc
     2221 tcaacacact ggcctccctg ggctacgccg tggttgtgat tgacggcagg ggctcctgtc
     2281 agcgagggct tcggttcgaa ggggccctga aaaaccaaat gggccaggtg gagatcgagg
     2341 accaggtgga gggcctgcag ttcgtggccg agaagtatgg cttcatcgac ctgagccgag
     2401 ttgccatcca tggctgggcc tacgggggct tcctctcgct catggggcta atccacaagc
     2461 cccaggtgtt caaggtggcc atcgcgggtg ccccggtcac cgtctggatg gcctacgaca
     2521 cagggtacac tgagcgctac atggacgtcc ctgagaacaa ccagcacggc tatgaggcgg
     2581 gttccgtggc cctgcacgtg gagaagctgc ccaatgagcc caaccgcttg cttatcctcc
     2641 acggcttcct ggacgaaaac gtgcactttt tccacacaaa cttcctcgtc tcccaactga
     2701 tccgagcagg gaaaccttac cagctccaga tctaccccaa cgagagacac agtattcgct
     2761 gccccgagtc gggcgagcac tatgaagtca cgttgctgca ctttctacag gaatacctct
     2821 agtctagagc ctgcagtctc gaggcatgcg gtaccaagct tgtcgagaag tactagagga
     2881 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     2941 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3001 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3061 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct
     3121 gatcactgat atcgcctagg agatccgaac cagataagtg aaatctagtt ccaaactatt
     3181 ttgtcatttt taattttcgt attagcttac gacgctacac ccagttccca tctattttgt
     3241 cactcttccc taaataatcc ttaaaaactc catttccacc cctcccagtt cccaactatt
     3301 ttgtccgccc acagcggggc atttttcttc ctgttatgtt tttaatcaaa catcctgcca
     3361 actccatgtg acaaaccgtc atcttcggct actttttctc tgtcacagaa tgaaaatttt
     3421 tctgtcatct cttcgttatt aatgtttgta attgactgaa tatcaacgct tatttgcagc
     3481 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3541 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3601 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3661 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3721 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3781 acgttcttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3841 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3901 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     3961 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4021 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4081 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4141 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4201 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4261 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4321 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4381 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4441 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4501 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4561 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4621 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     4681 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4741 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4801 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4861 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4921 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     4981 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5041 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5101 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5161 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5221 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5281 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5341 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5401 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5461 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5521 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5581 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5641 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5701 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctggcct
     5761 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5821 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     5881 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     5941 caccgcatag accagccgcg taacctggca aaatcggtta cggttgagta ataaatggat
     6001 gccctgcgta agcgggtgtg ggcggacaat aaagtcttaa actgaacaaa atagatctaa
     6061 actatgacaa taaagtctta aactagacag aatagttgta aactgaaatc agtccagtta
     6121 tgctgtgaaa aagcatactg gacttttgtt atggctaaag caaactcttc attttctgaa
     6181 gtgcaaattg cccgtcgtat taaagagggg cgtggccaag ggcatggtaa agactatatt
     6241 cgcggcgttg tgacaattta ccgaacaact ccgcggccgg gaagccgatc tcggcttgaa
     6301 cgaattgtta ggtggcggta cttgggtcga tatcaaagtg catcacttct tcccgtatgc
     6361 ccaactttgt atagagagcc actgcgggat cgtcaccgta atctgcttgc acgtagatca
     6421 cataagcacc aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg gtggcaatgc
     6481 cctgcctccg gtgctcgccg gagactgcga gatcatagat atagatctca ctacgcggct
     6541 gctcaaactt gggcagaacg taagccgcga gagcgccaac aaccgcttct tggtcgaagg
     6601 cagcaagcgc gatgaatgtc ttactacgga gcaagttccc gaggtaatcg gagtccggct
     6661 gatgttggga gtaggtggct acgtctccga actcacgacc gaaaagatca agagcagccc
     6721 gcatggattt gacttggtca gggccgagcc tacatgtgcg aatgatgccc atacttgagc
     6781 cacctaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctccataaca
     6841 tcaaacatcg acccacggcg taacgcgctt gctgcttgga tgcccgaggc atagactgta
     6901 caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt
     6961 cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta cttgcattac agtttacgaa
     7021 ccgaacaggc ttatgtcaac tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc
     7081 cggcaacctt gggcagcagc gaagtcgagg catttctgtc ctggctggcg aacgagcgca
     7141 aggtttcggt ctccacgcat cgtcaggcat tggcggcctt gctgttcttc tacggcaagg
     7201 tgctgtgcac ggatctgccc tggcttcagg agatcggaag acctcggccg tcgcggcgct
     7261 tgccggtggt gctgaccccg gatgaagtgg ttcgcatcct cggttttctg gaaggcgagc
     7321 atcgtttgtt cgcccaggac tctagctata gttctagtgg ttggctacgt atactccgga
     7381 atattaatag
//
