LOCUS       sequence-426378-        4367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for strep-tagged PCaspase (Prokaryotic 
            Caspase) R153A mutant from Haliangium ochraceum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4367)
  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 4367)
  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-PCaspase-R153A
FEATURES             Location/Qualifiers
     source          1..4367
                     /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"
     regulatory      367..376
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      2266..2293
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2336..2353)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2470..3015)
                     /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(2550..2569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3129..3223)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3254..4048)
                     /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(3365..3384)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3975..3994
                     /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 tagcggccat ggtggccgtg ttgttaacac cgggatcgtt aaagatctac cgggtggtga
      421 ctatcgtccc ggcgctcacc agttcctggt tccggaagac taccaaccga aagcacagac
      481 ctttaccggt atcctggact tcgagctgag gtatctagtt gcacgtctgg cagcacgtac
      541 cgaaaacgtt agcgttatcc tggattgttg tcatagcggt ggtgcaatag ctgcaccggg
      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 gcttctactg
     2161 ggcgccccgc ttcggcgggg tttttttgca agggatcggt tgtcgagtaa ggatctccag
     2221 gcatcctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt
     2281 gggcctttct gcgtttatag gatcctaact cgagcctagg gatatattcc gcttcctcgc
     2341 tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg
     2401 gcggagattt cctggaagat gccaggaaga tacttaacag ggaagtgaga gggccgcggc
     2461 aaagccgttt ttccataggc tccgcccccc tgacaagcat cacgaaatct gacgctcaaa
     2521 tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc
     2581 cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc
     2641 gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg
     2701 actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc
     2761 ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat
     2821 ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt
     2881 ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc
     2941 ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga
     3001 ccaaaacgat ctcaagaaga tcatcttatt aatcagataa aatatttcta gatttcagtg
     3061 caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgttacta
     3121 gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
     3181 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga gctctcgaac
     3241 cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
     3301 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
     3361 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
     3421 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     3481 cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca
     3541 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     3601 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     3661 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     3721 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     3781 cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca
     3841 gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct
     3901 tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc
     3961 cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac
     4021 ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag
     4081 atcatgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc
     4141 agggcttccc aaccttacca gagggcgccc cagctggcaa ttccgacgtc gaattcaaaa
     4201 gatcttaagt aagtaagagt atacgtatat cggctaataa cgtattaagg cgcttcggcg
     4261 ccttttttta tgggggtatt ttcatcccaa tccacacgtc caacgcacag caaacaccac
     4321 gtcgacccta tcagctgcgt gctttctatg agtcgttgct gcataac
//
