LOCUS       sequence-192693-        7086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminal flag-tagged pro-IL-1-beta in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7086)
  AUTHORS   Shi H, Wang Y, Li X, Zhan X, Tang M, Fina M, Su L, Pratt D, Bu CH, 
            Hildebrand S, Lyon S, Scott L, Quan J, Sun Q, Russell J, Arnett S, 
            Jurek P, Chen D, Kravchenko VV, Mathison JC, Moresco EM, Monson NL, 
            Ulevitch RJ, Beutler B
  TITLE     NLRP3 activation and mitosis are mutually exclusive events 
            coordinated by NEK7, a new inflammasome component.
  JOURNAL   Nat Immunol. 2016 Mar;17(3):250-8. doi: 10.1038/ni.3333. Epub 2015 
            Dec 7.
  PUBMED    26642356
REFERENCE   2  (bases 1 to 7086)
  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: "10.1038/ni.3333"; 
            journalName: "Nat Immunol"; date: "2016-03"; volume: "17"; issue: 
            "3"; pages: "250-8"
COMMENT     Sequence Label: pCMV-pro-Il1b-C-Flag
FEATURES             Location/Qualifiers
     source          1..7086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        178..557
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        558..761
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     712..732
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1629..1694
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1705..2327
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1904..1923)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2351..2371)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2369..2666
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2720..3514
                     /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(2774..2793)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3384..3403
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4166..4300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4203..4222)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4257..4276
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4305..4324)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4306..4324)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4338..4354)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4338..4354)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4351..4373)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4362..4378
                     /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(4386..4392,4393..4410,4411..4416))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4431..4452
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4569..4586)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4740..5328)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4820..4839)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5499..6290,6291..6359))
                     /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     6122..6141
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6360..6464)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6491..6946
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6578..6597)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6788..6809
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(7072..7086,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(7086,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca
       61 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
      121 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac
      181 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      241 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      301 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      361 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      421 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      481 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag
      541 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt
      601 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      661 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg
      721 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
      781 attaagcttg cggccgcgaa ttcatggcaa ctgttcctga actcaactgt gaaatgccac
      841 cttttgacag tgatgagaat gacctgttct ttgaagttga cggaccccaa aagatgaagg
      901 gctgcttcca aacctttgac ctgggctgtc ctgatgagag catccagctt caaatctcgc
      961 agcagcacat caacaagagc ttcaggcagg cagtatcact cattgtggct gtggagaagc
     1021 tgtggcagct acctgtgtct ttcccgtgga ccttccagga tgaggacatg agcaccttct
     1081 tttccttcat ctttgaagaa gagcccatcc tctgtgactc atgggatgat gatgataacc
     1141 tgctggtgtg tgacgttccc attagacaac tgcactacag gctccgagat aaacaacaaa
     1201 aaagcctcgt gctgtcggac ccatatgagc tgaaagctct ccacctcaat ggacagaata
     1261 tcaaccaaca agtgatattc tccatgagct ttgtacaagg agaaccaagc aacgacaaaa
     1321 tacctgtggc cttgggcctc aaaggaaaga atctatacct gtcctgtgta atgaaagacg
     1381 gcacacccac cctgcagctg gagagtgtgg atcccaagca atacccaaag aagaagatgg
     1441 aaaaacggtt tgtcttcaac aagatagaag tcaagagcaa agtggagttt gagtctgcag
     1501 agttccccaa ctggtacatc agcacctcac aagcagagca caagcctgtc ttcctgggaa
     1561 acaacagtgg tcaggacata attgacttca ccatggaatc cgtgtcttcc tctagaggat
     1621 cccgggctga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     1681 atgacgatga caagtagtga tcccgggtgg catccctgtg acccctcccc agtgcctctc
     1741 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     1801 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     1861 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     1921 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     1981 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     2041 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     2101 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     2161 actgctccct tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag
     2221 acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc
     2281 actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca
     2341 gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2401 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2461 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2521 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2581 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     2641 cagaagtagt gaggaggctt ttttggagga attgatcagc ttgggatctg atcaagagac
     2701 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     2761 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     2821 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     2881 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     2941 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3001 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3061 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3121 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3181 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3241 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     3301 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     3361 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     3421 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     3481 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     3541 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     3601 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     3661 ctcatgctgg agttcttcgc ccaccccggg ctcgatcccc tcgcgagttg gttcagctgc
     3721 tgcctgaggc tggacgacct cgcggagttc taccggcagt gcaaatccgt cggcatccag
     3781 gaaaccagca gcggctatcc gcgcatccat gcccccgaac tgcaggagtg gggaggcacg
     3841 atggccgctt tggtcgaccc ggacgggacg ctcctgcgcc tgatacagaa cgaattgctt
     3901 gcaggcatct catgagtgtg tcttcccgtt ttccgcctga ggtcactgcg tggatggagc
     3961 gctggcgcct gctgcgcgac ggcgagctgc tcaccaccca ctcgccaagc tggaaccgta
     4021 aaaaggccgc gttgctggcg tttttccata ggctccgccg atcataatca gccataccac
     4081 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     4141 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     4201 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4261 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aattccctat agtgagtcgt
     4321 attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4381 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4441 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4501 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4561 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4621 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4681 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4741 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4801 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4861 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4921 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4981 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5041 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5101 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5161 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5221 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5281 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5341 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5401 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5461 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5521 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5581 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5641 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5701 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5761 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5821 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5881 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5941 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6001 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6061 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6121 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6181 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6241 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6301 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6361 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6421 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6481 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     6541 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     6601 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     6661 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     6721 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     6781 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     6841 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     6901 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttccat tcgccattca
     6961 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     7021 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     7081 gacgtt
//
