LOCUS       sequence-423150-        5362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein overexpression in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5362)
  AUTHORS   Devant P, Dong Y, Mintseris J, Ma W, Gygi SP, Wu H, Kagan JC
  TITLE     Structural insights into cytokine cleavage by inflammatory 
            caspase-4.
  JOURNAL   Nature. 2023 Dec;624(7991):451-459. doi: 10.1038/s41586-023-06751-9.
            Epub 2023 Nov 22.
  PUBMED    37993712
REFERENCE   2  (bases 1 to 5362)
  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/s41586-023-06751-9"; journalName: "Nature"; date: 
            "2023-12"; volume: "624"; issue: "7991"; pages: "451-459"
COMMENT     Sequence Label: pFastBac_human-pro-IL-18
FEATURES             Location/Qualifiers
     source          1..5362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        482..586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(587..655,656..1447)
                     /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(805..824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2371..2388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2522..2746
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2813..3346)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3516..3521,3522..3538,3539..3544))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3896..3987
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3912..3929
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4054..4071
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4093..4113
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    4852..4986
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4889..4908)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4943..4962
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  5015..5180
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct ggccttttgc tcacatgttc
     2281 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2341 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2401 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catagaccag
     2461 ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa tggatgccct gcgtaagcgg
     2521 gtgtgggcgg acaataaagt cttaaactga acaaaataga tctaaactat gacaataaag
     2581 tcttaaacta gacagaatag ttgtaaactg aaatcagtcc agttatgctg tgaaaaagca
     2641 tactggactt ttgttatggc taaagcaaac tcttcatttt ctgaagtgca aattgcccgt
     2701 cgtattaaag aggggcgtgg ccaagggcat ggtaaagact atattcgcgg cgttgtgaca
     2761 atttaccgaa caactccgcg gccgggaagc cgatctcggc ttgaacgaat tgttaggtgg
     2821 cggtacttgg gtcgatatca aagtgcatca cttcttcccg tatgcccaac tttgtataga
     2881 gagccactgc gggatcgtca ccgtaatctg cttgcacgta gatcacataa gcaccaagcg
     2941 cgttggcctc atgcttgagg agattgatga gcgcggtggc aatgccctgc ctccggtgct
     3001 cgccggagac tgcgagatca tagatataga tctcactacg cggctgctca aacttgggca
     3061 gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca agcgcgatga
     3121 atgtcttact acggagcaag ttcccgaggt aatcggagtc cggctgatgt tgggagtagg
     3181 tggctacgtc tccgaactca cgaccgaaaa gatcaagagc agcccgcatg gatttgactt
     3241 ggtcagggcc gagcctacat gtgcgaatga tgcccatact tgagccacct aactttgttt
     3301 tagggcgact gccctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca
     3361 cggcgtaacg cgcttgctgc ttggatgccc gaggcataga ctgtacaaaa aaacagtcat
     3421 aacaagccat gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg
     3481 accagttgcg tgagcgcata cgctacttgc attacagttt acgaaccgaa caggcttatg
     3541 tcaactgggt tcgtgccttc atccgtttcc acggtgtgcg tcacccggca accttgggca
     3601 gcagcgaagt cgaggcattt ctgtcctggc tggcgaacga gcgcaaggtt tcggtctcca
     3661 cgcatcgtca ggcattggcg gccttgctgt tcttctacgg caaggtgctg tgcacggatc
     3721 tgccctggct tcaggagatc ggaagacctc ggccgtcgcg gcgcttgccg gtggtgctga
     3781 ccccggatga agtggttcgc atcctcggtt ttctggaagg cgagcatcgt ttgttcgccc
     3841 aggactctag ctatagttct agtggttggc tacgtatact ccggaatatt aatagatcat
     3901 ggagataatt aaaatgataa ccatctcgca aataaataag tattttactg ttttcgtaac
     3961 agttttgtaa taaaaaaacc tataaatatt ccggattatt cataccgtcc caccatcggg
     4021 cgcggatctc ggtccgaaac catgtcgtac taccatcacc atcaccatca cgattacgat
     4081 atcccaacga ccgaaaacct gtattttcag ggcgccatgg gatccgctgc tgaaccagta
     4141 gaagacaatt gcatcaactt tgtggcaatg aaatttattg acaatacgct ttactttata
     4201 gctgaagatg atgaaaacct ggaatcagat tactttggca agcttgaatc taaattatca
     4261 gtcataagaa atttgaatga ccaagttctc ttcattgacc aaggaaatcg gcctctattt
     4321 gaagatatga ctgattctga ctgtagagat aatgcacccc ggaccatatt tattataagt
     4381 atgtataaag atagccagcc tagaggtatg gctgtaacta tctctgtgaa gtgtgagaaa
     4441 atttcaactc tctcctgtga gaacaaaatt atttccttta aggaaatgaa tcctcctgat
     4501 aacatcaagg atacaaaaag tgacatcata ttctttcaga gaagtgtccc aggacatgat
     4561 aataagatgc aatttgaatc ttcatcatac gaaggatact ttctagcttg tgaaaaagag
     4621 agagaccttt ttaaactcat tttgaaaaaa gaggatgaat tgggggatag atctataatg
     4681 ttcactgttc aaaacgaaga ctagctcgag gcatgcggta ccaagcttgt cgagaagtac
     4741 tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     4801 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     4861 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4921 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4981 tggatctgat cactgatatc gcctaggaga tccgaaccag ataagtgaaa tctagttcca
     5041 aactattttg tcatttttaa ttttcgtatt agcttacgac gctacaccca gttcccatct
     5101 attttgtcac tcttccctaa ataatcctta aaaactccat ttccacccct cccagttccc
     5161 aactattttg tccgcccaca gcggggcatt tttcttcctg ttatgttttt aatcaaacat
     5221 cctgccaact ccatgtgaca aaccgtcatc ttcggctact ttttctctgt cacagaatga
     5281 aaatttttct gtcatctctt cgttattaat gtttgtaatt gactgaatat caacgcttat
     5341 ttgcagcctg aatggcgaat gg
//
