LOCUS       sequence-426379-        3053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for strep-tagged PC-sigma from 
            Haliangium ochraceum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3053)
  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 3053)
  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-PC-sigma
FEATURES             Location/Qualifiers
     source          1..3053
                     /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      1063..1090
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1133..1150)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1267..1812)
                     /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(1347..1366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1926..2020)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2051..2845)
                     /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(2162..2181)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2772..2791
                     /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 gagcgcggat catgatcacc gtgatgatga tgaaggtaaa cccgcactgg ctgaagatag
      301 cgaatacatt gatgagaacg ttctaggtga cgcagatgtt ctggaagatg atgatgaacg
      361 tggggaaagc gatggtgatg aagttagcga cgctgaactc tttgcaagct gggctgcagg
      421 tgataagcat gcagcagatg ttctgattcg tcgttacatg gataaactga gcggttactt
      481 tgctgcaaaa ctgtcgcagg gtgatagcgt tgcagacctg gttcagcaga ccctggaggg
      541 ttgccatcgt gcagcaccga gcttccgtgg tgaatgctcg tttcgtacct ttctgtacag
      601 ggttgcaggg aacaaactgc gtgactttct gcgtggtaag gcacgtaggc cccgtgcaga
      661 agacattgat gaaataagcg tagcagatct gaacccggga ccgagcacca ttcgtcgtca
      721 aaaccaccgt caggcagttc tgctggacgc actgcgtagc ctgccgctgc agatgcagga
      781 gctgctggaa ttccgttacc agcaagacct gagcatacgt gaaattgcag aaattctggg
      841 tattgaggtt agcgcaacca aaagccgtct gcgtcgtgca aaggaagcac tgcgtagggc
      901 actgggaact agcggtgctg atgcactgta gtgccgactc agttgctgct tctactgggc
      961 gccccgcttc ggcggggttt ttttgcaagg gatcggttgt cgagtaagga tctccaggca
     1021 tcctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg
     1081 cctttctgcg tttataggat cctaactcga gcctagggat atattccgct tcctcgctca
     1141 ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac gaacggggcg
     1201 gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg ccgcggcaaa
     1261 gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac gctcaaatca
     1321 gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gcggctccct
     1381 cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg
     1441 tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact
     1501 gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg
     1561 agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta
     1621 gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt
     1681 gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc
     1741 gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca
     1801 aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat ttcagtgcaa
     1861 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gttactagtg
     1921 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     1981 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct ctcgaacccc
     2041 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     2101 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     2161 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     2221 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     2281 catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt
     2341 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     2401 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     2461 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     2521 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     2581 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     2641 acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga
     2701 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     2761 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     2821 cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc
     2881 atgatcccct gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg
     2941 gcttcccaac cttaccagag ggcgccccag ctggcaattc cgacgtcgaa ttcaaaagat
     3001 cttaagtaag taagagtata cgtatatcgg ctaataacgt attaaggcgc ttc
//
