LOCUS       sequence-426423-        4576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for PCaspase from Haliangium ochraceum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4576)
  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 4576)
  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: pPCaspase
FEATURES             Location/Qualifiers
     source          1..4576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(33..79)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     94..116
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     promoter        join(123..128,129..146,147..153)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     regulatory      454..463
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2264..2282)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2278..2325
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(2590..3381,3382..3450))
                     /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     3213..3232
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3451..3555)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3629..4367)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
ORIGIN
        1 cgactcctgc attaggaaat gacaaacaac agataaaacg aaaggcccag tctttcgact
       61 gagcctttcg ttttatttga tgcctttaat taaagcggat aacaatttca cacaggaggc
      121 cgtttacact ttatgcttcc ggctcgtata atgtgtgggg aaaccacaac ggtttccctc
      181 tagaaataat tttgtttaac tataagaagg agatatacat atgatgaaaa gggctctgct
      241 gataggtgca ccgctggcac atagcgcaac ccatgcactg gcaggtgttg caaacgatat
      301 cgaccgtgca ggtgaaatcc tgcgtctgta tggttttcat tgtgatgacg ttctctttgg
      361 tgaacgtgca actagagatg gtatcgttgc acgtctggaa gcactgatcg ctgaaaccca
      421 ggctgatgat gctgttgtta tctattttag cggccatggt ggccgtgttg ttaacaccgg
      481 gatcgttaaa gatctaccgg gtggtgacta tcgtcccggc gctcaccagt tcctggttcc
      541 ggaagactac caaccgaaag cacagacctt taccggtatc ctggacttcg agctgaggta
      601 tctagttgca cgtctggcag cacgtaccga aaacgttagc gttatcctgg attgttgtca
      661 tagcggtggt gcaataagag caccgggtag accgaccccg aaagcactgg atgaaaaaag
      721 cctgaagccg catactgttc acctgaaccg tgttatccgt gaacgtctgg ttcaactgcg
      781 tgaatcgctg gcagctagcc gtgcaccgcg tctgacccag gatgctcatc cgcatctggt
      841 tcgtatcgaa gcagcacacg aattccgtct cgcatatgaa accaggatca acaaccgtca
      901 gcagggggtt ctgaccgcag catgggcaga caccctgtgg agataccgtg ggctggcagt
      961 tagctggcag gcactggcaa gcgagatcca ggcaagggca cgtgcatttg ttcccatgca
     1021 gtatgttcac atcgatggcc cgaccgaacg tcgtctgttc agcctggctc gtgagccggc
     1081 acgtcatagc ctggcaatcg ttcaggaaag ggacggtacc ctgtggctgg acggtggcag
     1141 atttcagggt gttgcaagcg gtgcacgttt cgcagttctg ccgccgacca gcaccgaaag
     1201 cgacatcgat agcgcactgg cagaagttcg tgcagttgaa gttatgccgg aacgtacccg
     1261 tgttgaactg gctaccgcag cagatgacat cccgagatgg caaggcctcc atgcatggcc
     1321 gctgcatgca cgtggcgctt ggaccggtgt agcagttcgt gctcagaccg cagcagtttg
     1381 tgctgcactc ctgagagcta tcgaagcaag ccagcgtatg catctgtgtc cggatagcga
     1441 cgggccggtt gctaccgtta tcgaactatc gagcgatgag ggcagcagga tcgcagttct
     1501 ggatgggcac ggtgtttgtg ttagcccccc ggttgcaacc cccgaacgtg cactcgcact
     1561 ggttgaaagg ctccagagag ctgcactcgt tcgtatgctg gggagcggta ccgatgagca
     1621 tgcactaccg atcccgctcg cactcagact ggaacgtgca gacggtatcg gtggtacctg
     1681 ccgtcatcta gcagaagggc gtaccgaacc gccgagcgaa ctgctactcg caagcgctgg
     1741 ggcacaggta catgcaatcg tagagaacca tgggagcgct atcgcaaccg atacccagga
     1801 tgtttatctg accatgttct gtatcgcagc agacggtcgt gttgaaagag ttagcaagca
     1861 gcagagcagc gggatccgtc tgtgtggggg tgcaaagcat aggctcgagt atgaactagg
     1921 tgcagggggt gcaagcctgc cggaatacag ggcagcagat tcgagcacca gcggctatgg
     1981 gctgcatagc ctcatagttg ttgcaaccga ccgtcagcag gagctgcgta acctggaaac
     2041 ctggtcgggt gaaggtagcg ctgcaaaact gcagccgggt gttctggatc tgaccgctag
     2101 caaacgtgca ccggctagac gtggctcggc tagcgcaagc gggccgctga gatatgctgt
     2161 tctacgtgta gatctgcgtg tagctgcagc agtagcaggg cgtgatgcac cgaccctgta
     2221 atgccgtggg acagcccgtc ttaattaacc taggctgctg ccaccgctga gcaataacta
     2281 gcataacccc ttggggcctc taaacgggtc ttgagtggtt ttttgctgaa acctcaggca
     2341 tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag
     2401 acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcatc gtggccggat
     2461 cttgcggccc ctcggcttga acgaattgtt agacagatta tcaaaaagga tcttcaccta
     2521 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2581 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2641 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2701 atctggcccc agtgctgcaa tgataccgcg ggacccacgc tcaccggctc cagatttatc
     2761 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2821 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2881 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2941 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3001 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3061 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3121 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3181 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3241 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3301 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3361 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3421 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3481 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3541 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gtagctagct cactcggtcg
     3601 ctacgctccg ggcgtgagac tgcggcgggc gctgcggaca catacaaagt tacccacaga
     3661 ttccgtggat aagcagggga ctaacatgtg aggcaaaaca gcagggccgc gccggtggcg
     3721 tttttccata ggctccgccc tcctgccaga gttcacataa acagacgctt ttccggtgca
     3781 tctgtgggag ccgtgaggct caaccatgaa tctgacagta cgggcgaaac ccgacaggac
     3841 ttaaagatcc ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg ttccgaccct
     3901 gccgtttacc ggatacctgt tccgcctttc tcccttacgg gaagtgtggc gctttctcat
     3961 agctcacaca ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg ggctgtaagc
     4021 aagaactccc cgttcagccc gactgctgcg ccttatccgg taactgttca cttgagtcca
     4081 acccggaaaa gcacggtaaa acgccactgg cagcagccat tggtaactgg gagttcgcag
     4141 aggatttgtt tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt ccggctacac
     4201 tggaaggaca gatttggttg ctgtgctctg cgaaagccag ttaccacggt taagcagttc
     4261 cccaactgac ttaaccttcg atcaaaccac ctccccaggt ggttttttcg tttacagggc
     4321 aaaagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctactgaac
     4381 cgctctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac
     4441 gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg agctgactgg
     4501 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     4561 attaattgcg ttgcgc
//
