LOCUS       sequence-426126-        3932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for strep-tagged SAVED-CHAT cA3-binding
            pocket mutant from Haliangium ochraceum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3932)
  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 3932)
  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-SAVED-CHAT-cbm
FEATURES             Location/Qualifiers
     source          1..3932
                     /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      1831..1858
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1901..1918)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2035..2580)
                     /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(2115..2134)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2694..2788)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2819..3613)
                     /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(2930..2949)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3540..3559
                     /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 agcgcgactc accatgctga tcatgactgc cagccgcaca gggagagcgt
      181 tcagccggat gaaatccagt gtatcctggt tctggacctg agcatcgata acgcaatcac
      241 cgcatgtagc gttaccccgc atctgccccg tgctgcacgt cgtgttgaac tacatctgaa
      301 cgactttggt gcagaacgtg caccgtacgg gggtgcaagc gatcgtcgta cctggcgttg
      361 ttggatgcag gcagttgatg caatgctggc tgacgcacgt gcacagctgg gtgcagaagt
      421 tgaatttact cattattatc tggcaggtcg tgcagcactc ccggtttttg catatctggg
      481 tctgagactg ggtgaagcag caaacatcac taccgttaac cgtcgtgatg acggttgttg
      541 ggatgttgtt ccgtgccagc gtccggcttc gagcgcagca tcgggggttt cgcccagcgc
      601 acgttttttt gacgaagttc gtggtctgga caccgatgag cgtagcagcg aaagcggtat
      661 ggttgcagtt tgggtaagca cccagcgtga tgttgatcgt gggctgctgc gtgcatttgc
      721 aagggcaaga ggtgaccgtg acctggcagg tatcgttagc ctgcgtgcac gtccggctgc
      781 aggtgatgac accggtgata tgcgtctcct ggaaggtgca gacggccccg atgctgcacg
      841 tgaactggtt aactgttttc gtagcatccc gaaccagtat ccgcgtagca gcggcctgat
      901 ggtttttgtt agcggcccgg ttaccctggc agcaatggtt gggcgtgcaa tcgcaccgga
      961 aatccatggc ccggtttggt ggccggcagc gcgtggtggg gagtacgagc cggcactgga
     1021 atatccgtgg ccgctgatca gcggcccccc gaggatcctg atcgcaaccg caaacgctcc
     1081 ggagggtgaa aacccgactc tggacgttga ggcagagctg aagcatctgg aagaagcact
     1141 ggcagaaccg cgtaaacgta aactgtgtga ggtacagcgt tgcccggcag caaccgtaag
     1201 cgatatcacc agcgcactgc gtagctttaa accgcatata ctgcatttta tcggtcatgg
     1261 tactgcactc ggtgtttatc tgaggagcgc agaacatgac ggtgcacagt ttgttcgtgg
     1321 tgaggatttc cagcagatga tcgcaacctc gctacgtcag aaggaccgtg aaatgcatct
     1381 ggttgttctg aacgcttgct gcacccacga actggcaaag gcactgaccg aacaggttag
     1441 ctgcaccatc ggtaccgata tcgaagttta tgattcggca agcatacatt ttgctgcacg
     1501 tttttatgat cacctggttc atggtaccag cgttcattat gcatttaacg cagcagttga
     1561 tgaatgtcgt gcacacagca ccagcggcca ggaagttttt tgcctgcacc cggcagcaga
     1621 acgtgcaccg gcaagcgcaa ccccgccggt acgtgcagat gaactggttt tctttagccc
     1681 ggcagaacgt catagctaat gccgactcag ttgctgcttc tactgggcgc cccgcttcgg
     1741 cggggttttt ttgcaaggga tcggttgtcg agtaaggatc tccaggcatc ctgttgtttg
     1801 tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt
     1861 tataggatcc taactcgagc ctagggatat attccgcttc ctcgctcact gactcgctac
     1921 gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg
     1981 aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca
     2041 taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa
     2101 cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc
     2161 tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc
     2221 cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac
     2281 cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2341 aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc
     2401 ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct
     2461 ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc
     2521 tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa
     2581 gaagatcatc ttattaatca gataaaatat ttctagattt cagtgcaatt tatctcttca
     2641 aatgtagcac ctgaagtcag ccccatacga tataagttgt tactagtgct tggattctca
     2701 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     2761 gtcattactg gatctatcaa caggagtcca agcgagctct cgaaccccag agtcccgctc
     2821 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
     2881 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
     2941 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc
     3001 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga
     3061 cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga
     3121 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
     3181 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
     3241 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
     3301 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
     3361 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
     3421 ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
     3481 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
     3541 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
     3601 cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatcat gatcccctgc
     3661 gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct
     3721 taccagaggg cgccccagct ggcaattccg acgtcgaatt caaaagatct taagtaagta
     3781 agagtatacg tatatcggct aataacgtat taaggcgctt cggcgccttt ttttatgggg
     3841 gtattttcat cccaatccac acgtccaacg cacagcaaac accacgtcga ccctatcagc
     3901 tgcgtgcttt ctatgagtcg ttgctgcata ac
//
