LOCUS       sequence-328370-        8098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression vector for FLAG-tagged DPP9S. LL100EE 
            mutant is loss of CARD8 and NLRP1 binding..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8098)
  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 8098)
  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: pcDNA3.1 LIC 6A FLAG-TEV-DPP9S LL100EE
FEATURES             Location/Qualifiers
     source          1..8098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      929..938
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             944..967
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             989..1009
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     promoter        complement(3651..3669)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3652..3669)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3689..3706)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3695..3919
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3965..4393
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4052..4071)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4262..4283
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4402..4422)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4407..4736
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4587..4722
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4649..4668
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4803..5597
                     /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(4857..4876)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5467..5486
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5771..5892
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5808..5827)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5862..5881
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5941..5957)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5941..5957)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5954..5976)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5965..5981
                     /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(join(5989..5995,5996..6013,6014..6019))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6034..6055
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6172..6189)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6343..6931)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6423..6442)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7102..7893,7894..7962))
                     /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     7725..7744
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7963..8067)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccctacttcc aatccaatgc caccatggat atcgactaca aggacgatga
      961 cgacaaggat tacgatatcc caacgaccga aaacctgtat tttcagggcg ccatgggatc
     1021 catggccacc accgggaccc caacggccga ccgaggcgac gcagccgcca cagatgaccc
     1081 ggccgcccgc ttccaggtgc agaagcactc gtgggacggg ctccggagca tcatccacgg
     1141 cagccgcaag tactcgggcc tcattgtcaa caaggcgccc cacgacttcc agtttgtgca
     1201 gaagacggat gagtctgggc cccactccca ccgcctctac tacctgggaa tgccatatgg
     1261 cagccgagag aactccctcc tctactctga gattcccaag aaggtccgga aagaggctga
     1321 agaactcctg tcctggaagc agatgctgga tcatttccag gccacgcccc accatggggt
     1381 ctactctcgg gaggaggagc tgctgaggga gcggaaacgc ctgggggtct tcggcatcac
     1441 ctcctacgac ttccacagcg agagtggcct cttcctcttc caggccagca acagcctctt
     1501 ccactgccgc gacggcggca agaacggctt catggtgtcc cctatgaaac cgctggaaat
     1561 caagacccag tgctcagggc cccggatgga ccccaaaatc tgccctgccg accctgcctt
     1621 cttctccttc atcaataaca gcgacctgtg ggtggccaac atcgagacag gcgaggagcg
     1681 gcggctgacc ttctgccacc aaggtttatc caatgtcctg gatgacccca agtctgcggg
     1741 tgtggccacc ttcgtcatac aggaagagtt cgaccgcttc actgggtact ggtggtgccc
     1801 cacagcctcc tgggaaggtt cagagggcct caagacgctg cgaatcctgt atgaggaagt
     1861 cgatgagtcc gaggtggagg tcattcacgt cccctctcct gcgctagaag aaaggaagac
     1921 ggactcgtat cggtacccca ggacaggcag caagaatccc aagattgcct tgaaactggc
     1981 tgagttccag actgacagcc agggcaagat cgtctcgacc caggagaagg agctggtgca
     2041 gcccttcagc tcgctgttcc cgaaggtgga gtacatcgcc agggccgggt ggacccggga
     2101 tggcaaatac gcctgggcca tgttcctgga ccggccccag cagtggctcc agctcgtcct
     2161 cctccccccg gccctgttca tcccgagcac agagaatgag gagcagcggc tagcctctgc
     2221 cagagctgtc cccaggaatg tccagccgta tgtggtgtac gaggaggtca ccaacgtctg
     2281 gatcaatgtt catgacatct tctatccctt cccccaatca gagggagagg acgagctctg
     2341 ctttctccgc gccaatgaat gcaagaccgg cttctgccat ttgtacaaag tcaccgccgt
     2401 tttaaaatcc cagggctacg attggagtga gcccttcagc cccggggaag atgaatttaa
     2461 gtgccccatt aaggaagaga ttgctctgac cagcggtgaa tgggaggttt tggcgaggca
     2521 cggctccaag atctgggtca atgaggagac caagctggtg tacttccagg gcaccaagga
     2581 cacgccgctg gagcaccacc tctacgtggt cagctatgag gcggccggcg agatcgtacg
     2641 cctcaccacg cccggcttct cccatagctg ctccatgagc cagaacttcg acatgttcgt
     2701 cagccactac agcagcgtga gcacgccgcc ctgcgtgcac gtctacaagc tgagcggccc
     2761 cgacgacgac cccctgcaca agcagccccg cttctgggct agcatgatgg aggcagccag
     2821 ctgccccccg gattatgttc ctccagagat cttccatttc cacacgcgct cggatgtgcg
     2881 gctctacggc atgatctaca agccccacgc cttgcagcca gggaagaagc accccaccgt
     2941 cctctttgta tatggaggcc cccaggtgca gctggtgaat aactccttca aaggcatcaa
     3001 gtacttgcgg ctcaacacac tggcctccct gggctacgcc gtggttgtga ttgacggcag
     3061 gggctcctgt cagcgagggc ttcggttcga aggggccctg aaaaaccaaa tgggccaggt
     3121 ggagatcgag gaccaggtgg agggcctgca gttcgtggcc gagaagtatg gcttcatcga
     3181 cctgagccga gttgccatcc atggctggtc ctacgggggc ttcctctcgc tcatggggct
     3241 aatccacaag ccccaggtgt tcaaggtggc catcgcgggt gccccggtca ccgtctggat
     3301 ggcctacgac acagggtaca ctgagcgcta catggacgtc cctgagaaca accagcacgg
     3361 ctatgaggcg ggttccgtgg ccctgcacgt ggagaagctg cccaatgagc ccaaccgctt
     3421 gcttatcctc cacggcttcc tggacgaaaa cgtgcacttt ttccacacaa acttcctcgt
     3481 ctcccaactg atccgagcag ggaaacctta ccagctccag atctacccca acgagagaca
     3541 cagtattcgc tgccccgagt cgggcgagca ctatgaagtc acgttgctgc actttctaca
     3601 ggaatacctc tagtaataac attggaagtg gataatctag agggccctat tctatagtgt
     3661 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3721 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3781 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3841 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3901 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3961 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4021 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4081 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4141 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4201 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4261 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4321 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4381 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4441 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4501 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4561 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4621 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4681 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4741 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4801 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4861 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4921 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4981 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5041 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5101 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5161 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5221 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5281 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5341 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5401 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5461 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5521 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5581 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5641 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5701 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5761 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5821 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5881 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5941 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6001 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6061 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6121 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6181 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6241 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6301 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6361 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6421 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6481 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6541 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6601 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6661 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6721 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6781 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6841 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6901 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6961 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7021 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7081 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7141 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7201 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7261 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7321 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7381 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7441 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7501 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7561 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7621 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7681 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7741 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7801 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7861 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7921 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcatta
     7981 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8041 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
