LOCUS       sequence-440342-        6057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SAVED-CHAT cA3 binding pocket 
            mutant and PCaspase from Haliangium ochraceum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6057)
  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 6057)
  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: pSAVED-CHAT-cbm_PCaspase
FEATURES             Location/Qualifiers
     source          1..6057
                     /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      2052..2061
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3862..3880)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3876..3923
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(4071..4862,4863..4931))
                     /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     4694..4713
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4932..5036)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5110..5848)
                     /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 atgactcacc atgctgatca
      241 tgactgccag ccgcacaggg agagcgttca gccggatgaa atccagtgta tcctggttct
      301 ggacctgagc atcgataacg caatcaccgc atgtagcgtt accccgcatc tgccccgtgc
      361 tgcacgtcgt gttgaactac atctgaacga ctttggtgca gaacgtgcac cgtacggggg
      421 tgcaagcgat cgtcgtacct ggcgttgttg gatgcaggca gttgatgcaa tgctggctga
      481 cgcacgtgca cagctgggtg cagaagttga atttactcat tattatctgg caggtcgtgc
      541 agcactcccg gtttttgcat atctgggtct gagactgggt gaagcagcaa acatcactac
      601 cgttaaccgt cgtgatgacg gttgttggga tgttgttccg tgccagcgtc cggcttcgag
      661 cgcagcatcg ggggtttcgc ccagcgcacg tttttttgac gaagttcgtg gtctggacac
      721 cgatgagcgt agcagcgaaa gcggtatggt tgcagtttgg gtaagcaccc agcgtgatgt
      781 tgatcgtggg ctgctgcgtg catttgcaag ggcaagaggt gaccgtgacc tggcaggtat
      841 cgttagcctg cgtgcacgtc cggctgcagg tgatgacacc ggtgatatgc gtctcctgga
      901 aggtgcagac ggccccgatg ctgcacgtga actggttaac tgttttcgta gcatcccgaa
      961 ccagtatccg cgtagcagcg gcctgatggt ttttgttagc ggcccggtta ccctggcagc
     1021 aatggttggg cgtgcaatcg caccggaaat ccatggcccg gtttggtggc cggcagcgcg
     1081 tggtggggag tacgagccgg cactggaata tccgtggccg ctgatcagcg gccccccgag
     1141 gatcctgatc gcaaccgcaa acgctccgga gggtgaaaac ccgactctgg acgttgaggc
     1201 agagctgaag catctggaag aagcactggc agaaccgcgt aaacgtaaac tgtgtgaggt
     1261 acagcgttgc ccggcagcaa ccgtaagcga tatcaccagc gcactgcgta gctttaaacc
     1321 gcatatactg cattttatcg gtcatggtac tgcactcggt gtttatctga ggagcgcaga
     1381 acatgacggt gcacagtttg ttcgtggtga ggatttccag cagatgatcg caacctcgct
     1441 acgtcagaag gaccgtgaaa tgcatctggt tgttctgaac gcttgctgca cccacgaact
     1501 ggcaaaggca ctgaccgaac aggttagctg caccatcggt accgatatcg aagtttatga
     1561 ttcggcaagc atacattttg ctgcacgttt ttatgatcac ctggttcatg gtaccagcgt
     1621 tcattatgca tttaacgcag cagttgatga atgtcgtgca cacagcacca gcggccagga
     1681 agttttttgc ctgcacccgg cagcagaacg tgcaccggca agcgcaaccc cgccggtacg
     1741 tgcagatgaa ctggttttct ttagcccggc agaacgtcat agctaataac atacgcgtaa
     1801 caaggtaagg acaaattaat gatgaaaagg gctctgctga taggtgcacc gctggcacat
     1861 agcgcaaccc atgcactggc aggtgttgca aacgatatcg accgtgcagg tgaaatcctg
     1921 cgtctgtatg gttttcattg tgatgacgtt ctctttggtg aacgtgcaac tagagatggt
     1981 atcgttgcac gtctggaagc actgatcgct gaaacccagg ctgatgatgc tgttgttatc
     2041 tattttagcg gccatggtgg ccgtgttgtt aacaccggga tcgttaaaga tctaccgggt
     2101 ggtgactatc gtcccggcgc tcaccagttc ctggttccgg aagactacca accgaaagca
     2161 cagaccttta ccggtatcct ggacttcgag ctgaggtatc tagttgcacg tctggcagca
     2221 cgtaccgaaa acgttagcgt tatcctggat tgttgtcata gcggtggtgc aataagagca
     2281 ccgggtagac cgaccccgaa agcactggat gaaaaaagcc tgaagccgca tactgttcac
     2341 ctgaaccgtg ttatccgtga acgtctggtt caactgcgtg aatcgctggc agctagccgt
     2401 gcaccgcgtc tgacccagga tgctcatccg catctggttc gtatcgaagc agcacacgaa
     2461 ttccgtctcg catatgaaac caggatcaac aaccgtcagc agggggttct gaccgcagca
     2521 tgggcagaca ccctgtggag ataccgtggg ctggcagtta gctggcaggc actggcaagc
     2581 gagatccagg caagggcacg tgcatttgtt cccatgcagt atgttcacat cgatggcccg
     2641 accgaacgtc gtctgttcag cctggctcgt gagccggcac gtcatagcct ggcaatcgtt
     2701 caggaaaggg acggtaccct gtggctggac ggtggcagat ttcagggtgt tgcaagcggt
     2761 gcacgtttcg cagttctgcc gccgaccagc accgaaagcg acatcgatag cgcactggca
     2821 gaagttcgtg cagttgaagt tatgccggaa cgtacccgtg ttgaactggc taccgcagca
     2881 gatgacatcc cgagatggca aggcctccat gcatggccgc tgcatgcacg tggcgcttgg
     2941 accggtgtag cagttcgtgc tcagaccgca gcagtttgtg ctgcactcct gagagctatc
     3001 gaagcaagcc agcgtatgca tctgtgtccg gatagcgacg ggccggttgc taccgttatc
     3061 gaactatcga gcgatgaggg cagcaggatc gcagttctgg atgggcacgg tgtttgtgtt
     3121 agccccccgg ttgcaacccc cgaacgtgca ctcgcactgg ttgaaaggct ccagagagct
     3181 gcactcgttc gtatgctggg gagcggtacc gatgagcatg cactaccgat cccgctcgca
     3241 ctcagactgg aacgtgcaga cggtatcggt ggtacctgcc gtcatctagc agaagggcgt
     3301 accgaaccgc cgagcgaact gctactcgca agcgctgggg cacaggtaca tgcaatcgta
     3361 gagaaccatg ggagcgctat cgcaaccgat acccaggatg tttatctgac catgttctgt
     3421 atcgcagcag acggtcgtgt tgaaagagtt agcaagcagc agagcagcgg gatccgtctg
     3481 tgtgggggtg caaagcatag gctcgagtat gaactaggtg cagggggtgc aagcctgccg
     3541 gaatacaggg cagcagattc gagcaccagc ggctatgggc tgcatagcct catagttgtt
     3601 gcaaccgacc gtcagcagga gctgcgtaac ctggaaacct ggtcgggtga aggtagcgct
     3661 gcaaaactgc agccgggtgt tctggatctg accgctagca aacgtgcacc ggctagacgt
     3721 ggctcggcta gcgcaagcgg gccgctgaga tatgctgttc tacgtgtaga tctgcgtgta
     3781 gctgcagcag tagcagggcg tgatgcaccg accctgtaat gccgtgggac agcccgtctt
     3841 aattaaccta ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     3901 aacgggtctt gaggggtttt ttggggtcat cgtggccgga tcttgcggcc cctcggcttg
     3961 aacgaattgt tagacagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4021 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4081 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4141 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4201 atgataccgc gggacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4261 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4321 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4381 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4441 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4501 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4561 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4621 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4681 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4741 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4801 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4861 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4921 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4981 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5041 tttccccgaa aagtgccacc tgtagctagc tcactcggtc gctacgctcc gggcgtgaga
     5101 ctgcggcggg cgctgcggac acatacaaag ttacccacag attccgtgga taagcagggg
     5161 actaacatgt gaggcaaaac agcagggccg cgccggtggc gtttttccat aggctccgcc
     5221 ctcctgccag agttcacata aacagacgct tttccggtgc atctgtggga gccgtgaggc
     5281 tcaaccatga atctgacagt acgggcgaaa cccgacagga cttaaagatc cccaccgttt
     5341 ccggcgggtc gctccctctt gcgctctcct gttccgaccc tgccgtttac cggatacctg
     5401 ttccgccttt ctcccttacg ggaagtgtgg cgctttctca tagctcacac actggtatct
     5461 cggctcggtg taggtcgttc gctccaagct gggctgtaag caagaactcc ccgttcagcc
     5521 cgactgctgc gccttatccg gtaactgttc acttgagtcc aacccggaaa agcacggtaa
     5581 aacgccactg gcagcagcca ttggtaactg ggagttcgca gaggatttgt ttagctaaac
     5641 acgcggttgc tcttgaagtg tgcgccaaag tccggctaca ctggaaggac agatttggtt
     5701 gctgtgctct gcgaaagcca gttaccacgg ttaagcagtt ccccaactga cttaaccttc
     5761 gatcaaacca cctccccagg tggttttttc gtttacaggg caaaagatta cgcgcagaaa
     5821 aaaaggatct caagaagatc ctttgatctt ttctactgaa ccgctctaga tttcagtgca
     5881 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     5941 atgttagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     6001 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgc
//
