LOCUS       sequence-426375-        4415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for strep-tagged catalytically dead 
            PCaspase (Prokaryotic Caspase) from Haliangium ochraceum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4415)
  AUTHORS   Steens JA, Bravo JPK, Salazar CRP, Yildiz C, Amieiro AM, Kostlbacher
            S, Prinsen SHP, Andres AS, Patinios C, Bardis A, Barendregt A, 
            Scheltema RA, Ettema TJG, van der Oost J, Taylor DW, Staals RHJ
  TITLE     Type III-B CRISPR-Cas cascade of proteolytic cleavages.
  JOURNAL   Science. 2024 Feb 2;383(6682):512-519. doi: 10.1126/science.adk0378.
            Epub 2024 Feb 1.
  PUBMED    38301007
REFERENCE   2  (bases 1 to 4415)
  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.adk0378"; journalName: "Science"; date: "2024-02-2-
            2"; volume: "383"; issue: "6682"; pages: "512-519"
COMMENT     Sequence Label: pJS-BCD-Strep-dPCaspase
FEATURES             Location/Qualifiers
     source          1..4415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             107..130
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      2314..2341
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2384..2401)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2518..3063)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2598..2617)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3177..3271)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3302..4096)
                     /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(3413..3432)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4023..4042
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 taatacgact cactataggg gcccaagttc acttaaaaag gagatcaaca atgaaagcaa
       61 ttttcgtact gaaacatctt aatcatgcag gggagggttt ctaatgtgga gccacccgca
      121 gttcgaaaaa agcgcgatga aaagggctct gctgataggt gcaccgctgg cacatagcgc
      181 aacccatgca ctggcaggtg ttgcaaacga tatcgaccgt gcaggtgaaa tcctgcgtct
      241 gtatggtttt cattgtgatg acgttctctt tggtgaacgt gcaactagag atggtatcgt
      301 tgcacgtctg gaagcactga tcgctgaaac ccaggctgat gatgctgttg ttatctattt
      361 tagcggcgct ggtggccgtg ttgttaacac cgggatcgtt aaagatctac cgggtggtga
      421 ctatcgtccc ggcgctcacc agttcctggt tccggaagac taccaaccga aagcacagac
      481 ctttaccggt atcctggact tcgagctgag gtatctagtt gcacgtctgg cagcacgtac
      541 cgaaaacgtt agcgttatcc tggattgtgc tcatagcggt ggtgcaataa gagcaccggg
      601 tagaccgacc ccgaaagcac tggatgaaaa aagcctgaag ccgcatactg ttcacctgaa
      661 ccgtgttatc cgtgaacgtc tggttcaact gcgtgaatcg ctggcagcta gccgtgcacc
      721 gcgtctgacc caggatgctc atccgcatct ggttcgtatc gaagcagcac acgaattccg
      781 tctcgcatat gaaaccagga tcaacaaccg tcagcagggg gttctgaccg cagcatgggc
      841 agacaccctg tggagatacc gtgggctggc agttagctgg caggcactgg caagcgagat
      901 ccaggcaagg gcacgtgcat ttgttcccat gcagtatgtt cacatcgatg gcccgaccga
      961 acgtcgtctg ttcagcctgg ctcgtgagcc ggcacgtcat agcctggcaa tcgttcagga
     1021 aagggacggt accctgtggc tggacggtgg cagatttcag ggtgttgcaa gcggtgcacg
     1081 tttcgcagtt ctgccgccga ccagcaccga aagcgacatc gatagcgcac tggcagaagt
     1141 tcgtgcagtt gaagttatgc cggaacgtac ccgtgttgaa ctggctaccg cagcagatga
     1201 catcccgaga tggcaaggcc tccatgcatg gccgctgcat gcacgtggcg cttggaccgg
     1261 tgtagcagtt cgtgctcaga ccgcagcagt ttgtgctgca ctcctgagag ctatcgaagc
     1321 aagccagcgt atgcatctgt gtccggatag cgacgggccg gttgctaccg ttatcgaact
     1381 atcgagcgat gagggcagca ggatcgcagt tctggatggg cacggtgttt gtgttagccc
     1441 cccggttgca acccccgaac gtgcactcgc actggttgaa aggctccaga gagctgcact
     1501 cgttcgtatg ctggggagcg gtaccgatga gcatgcacta ccgatcccgc tcgcactcag
     1561 actggaacgt gcagacggta tcggtggtac ctgccgtcat ctagcagaag ggcgtaccga
     1621 accgccgagc gaactgctac tcgcaagcgc tggggcacag gtacatgcaa tcgtagagaa
     1681 ccatgggagc gctatcgcaa ccgataccca ggatgtttat ctgaccatgt tctgtatcgc
     1741 agcagacggt cgtgttgaaa gagttagcaa gcagcagagc agcgggatcc gtctgtgtgg
     1801 gggtgcaaag cataggctcg agtatgaact aggtgcaggg ggtgcaagcc tgccggaata
     1861 cagggcagca gattcgagca ccagcggcta tgggctgcat agcctcatag ttgttgcaac
     1921 cgaccgtcag caggagctgc gtaacctgga aacctggtcg ggtgaaggta gcgctgcaaa
     1981 actgcagccg ggtgttctgg atctgaccgc tagcaaacgt gcaccggcta gacgtggctc
     2041 ggctagcgca agcgggccgc tgagatatgc tgttctacgt gtagatctgc gtgtagctgc
     2101 agcagtagca gggcgtgatg caccgaccct gtaatgccga ctcagttgct gtctcatgcc
     2161 gactcagttg ctgtcggtct catgccgact cagttgctgc ttctactggg cgccccgctt
     2221 cggcggggtt tttttgcaag ggatcggttg tcgagtaagg atctccaggc atcctgttgt
     2281 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
     2341 gtttatagga tcctaactcg agcctaggga tatattccgc ttcctcgctc actgactcgc
     2401 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     2461 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     2521 ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     2581 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     2641 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     2701 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     2761 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2821 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     2881 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     2941 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg
     3001 ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct
     3061 caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca atttatctct
     3121 tcaaatgtag cacctgaagt cagccccata cgatataagt tgttactagt gcttggattc
     3181 tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct
     3241 gaggtcatta ctggatctat caacaggagt ccaagcgagc tctcgaaccc cagagtcccg
     3301 ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga
     3361 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac
     3421 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga
     3481 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca
     3541 cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg
     3601 cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag
     3661 tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa
     3721 gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt
     3781 gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt
     3841 cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc
     3901 gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa
     3961 ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt
     4021 gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc
     4081 catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcagat catgatcccc
     4141 tgcgccatca gatccttggc ggcaagaaag ccatccagtt tactttgcag ggcttcccaa
     4201 ccttaccaga gggcgcccca gctggcaatt ccgacgtcga attcaaaaga tcttaagtaa
     4261 gtaagagtat acgtatatcg gctaataacg tattaaggcg cttcggcgcc tttttttatg
     4321 ggggtatttt catcccaatc cacacgtcca acgcacagca aacaccacgt cgaccctatc
     4381 agctgcgtgc tttctatgag tcgttgctgc ataac
//
