LOCUS       sequence-426380-        2810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for strep-tagged Pci from Haliangium 
            ochraceum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2810)
  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 2810)
  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-PCi
FEATURES             Location/Qualifiers
     source          1..2810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..131
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        112..130
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             218..241
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      820..847
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(890..907)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1024..1569)
                     /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(1104..1123)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1683..1777)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1808..2602)
                     /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(1919..1938)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2529..2548
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ggcgcctttt tttatggggg tattttcatc ccaatccaca cgtccaacgc acagcaaaca
       61 ccacgtcgac cctatcagct gcgtgctttc tatgagtcgt tgctgcataa ctaatacgac
      121 tcactatagg ggcccaagtt cacttaaaaa ggagatcaac aatgaaagca attttcgtac
      181 tgaaacatct taatcatgca ggggagggtt tctaatgtgg agccacccgc agttcgagaa
      241 gagcgcgcac aggattgacg tttcgggtcc gctgtacctg gattgtcaac agagcgggga
      301 caccgttcag ctgattcacc aaaccaccgt tctggttgaa ccggcaattg aactatacgg
      361 gaccgatacc agcaccaccc tggaaattcg tgactttgaa gatgaaaacg gaagatgtcg
      421 tgttaacgtt tttaccattg aacgtgacgg taccctgacc ccgtgggaac gtctagcaag
      481 cggagcagtt tcgccggcag tttggagcat tattgagcgt ggaggggagc gtgctgctct
      541 gctggttccg gttccgtacg caccgctggg tgaactgacc gcagacctgg ttgatcaatt
      601 tgcaccgcag gattacaccc cgtcgagggt tagaattgtt ctcccgccgc cggaagacga
      661 taaaccgccg ccgcacgatg accaataatg ccgactcagt tgctgcttct actgggcgcc
      721 ccgcttcggc ggggtttttt tgcaagggat cggttgtcga gtaaggatct ccaggcatcc
      781 tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct
      841 ttctgcgttt ataggatcct aactcgagcc tagggatata ttccgcttcc tcgctcactg
      901 actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag
      961 atttcctgga agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc
     1021 gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg
     1081 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt
     1141 gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt
     1201 gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta
     1261 tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1321 ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag
     1381 gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac
     1441 tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa
     1501 aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa
     1561 cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc agtgcaattt
     1621 atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgtt actagtgctt
     1681 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
     1741 agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctctc gaaccccaga
     1801 gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag
     1861 cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa
     1921 tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt
     1981 cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat
     2041 gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg
     2101 ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca
     2161 tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg
     2221 gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag
     2281 caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc
     2341 ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg
     2401 atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
     2461 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
     2521 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
     2581 gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagatcatg
     2641 atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact ttgcagggct
     2701 tcccaacctt accagagggc gccccagctg gcaattccga cgtcgaattc aaaagatctt
     2761 aagtaagtaa gagtatacgt atatcggcta ataacgtatt aaggcgcttc
//
