LOCUS       sequence-418339-        2885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pDonor_ShCAST_kanR with modification of resistance cassette from KmR
            to TpR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2885)
  AUTHORS   Yap ZL, Rahman ASMZ, Hogan AM, Levin DB, Cardona ST
  TITLE     A CRISPR-Cas-associated transposon system for genome editing in 
            Burkholderia cepacia complex species.
  JOURNAL   Appl Environ Microbiol. 2024 Jul 24;90(7):e0069924. doi: 
            10.1128/aem.00699-24. Epub 2024 Jun 13.
  PUBMED    38869300
REFERENCE   2  (bases 1 to 2885)
  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"; journalName: "Appl 
            Environ Microbiol."; date: "2024-07-24"; volume: "90(7):e0069924. 
            doi"; pages: " 10.1128/aem.00699-24. Epub 2024 Jun 13"
COMMENT     Sequence Label: pDonorTp
FEATURES             Location/Qualifiers
     source          1..2885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(395..442)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     terminator      complement(490..576)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(679..773)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1117..1473
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            1524..1632
                     /note="incP origin of transfer"
     protein_bind    complement(2075..2122)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        join(2518..2523,2524..2540,2541..2546)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(2872..2885,1..223)
                     /codon_start=1
                     /gene="dhfrIIb from plasmid R388"
                     /product="E. coli plasmid-associated dihydrofolate 
                     reductase"
                     /label=TpR
                     /note="confers resistance to trimethoprim"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
ORIGIN
        1 cgatgaagcc aacgctcccg ttgcagggca gtttgcgctt cccctgagtg ccacctttgg
       61 cttaggggat cgcgtacgca agaaatctgg tgccgcttgg cagggtcaag tcgtcggttg
      121 gtattgcaca aaactcactc ctgaaggcta tgcggtcgag tccgaatccc acccaggctc
      181 agtgcaaatt tatcctgtgg ctgcacttga acgtgtggcc taagcgagct cttaattaat
      241 ttaaatctag actagtgcgg ccgcacttgt gtataagagt cagaattcta ctcgagggaa
      301 gagtatgagg atccaacatt tccaatcact agtgaattat ctagaattat tccattgagt
      361 aagtttttaa gcacatcagc ttcaaaagcg ctctgaagtt cctatacttt ctagagaata
      421 ggaacttcgg aataggtact tcaagatccc caattcgaga tcgtccgggc cgcaagctcc
      481 tagcggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
      541 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctcaagctag agagtcatta
      601 ccccaggcgt ttaagggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact
      661 catcgcagtc tagcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct
      721 gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagctc
      781 agctaattaa ggcgacagtc aatttgtcat tatgaaaata cacaaaagct ttttcctatc
      841 ttgcaaagcg acagctaatt tgtcacaatc acggacaacg acatctattt tgtcactgca
      901 aagaggttat gctaaaactg ccaaagcgct ataatctata ctgtataagg attttactga
      961 tgacaataat ttgtcacaac gacatataat tagtcactgt acacgtagag acgtagcaat
     1021 gctacctctc tacaatggtt ttgtatggtg cactctcagt acaatcttgt agcaccgcct
     1081 acatacctcg ctctgctaat cctgttacca gccggttgtc agccgttaag tgttcctgtg
     1141 tcactcaaaa ttgctttgag aggctctaag ggcttctcag tgcgttacat ccctggcttg
     1201 ttgtccacaa ccgttaaacc ttaaaagctt taaaagcctt atatattctt ttttttctta
     1261 taaaacttaa aaccttagag gctatttaag ttgctgattt atattaattt tattgttcaa
     1321 acatgagagc ttagtacgtg aaacatgaga gcttagtacg ttagccatga gagcttagta
     1381 cgttagccat gagggtttag ttcgttaaac atgagagctt agtacgttaa acttgagagc
     1441 ttagtacgtg aaacatgaga gcttagtacg tactatcaac aggttgaact gcccatgttc
     1501 tttcctgcgt tatcagagct tatcggccag cctcgcagag caggattccc gttgagcacc
     1561 gccaggtgcg aataagggac agtgaagaag gaacacccgc tcgcgggtgg gcctacttca
     1621 cctatcctgc ccggctgacg ccgttggata caccaaggaa agtctacacg aaccctttgg
     1681 caaaatcctg tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc tataatgacc
     1741 ccgaagcagg gttatgcagc ggaaagtaaa aaatttttag tttattagac atctccacaa
     1801 aaggcgtagt gtacagtgac aaattatctg tcgtcggtga cagattaatg tcattgtgac
     1861 tatttaattg tcgtcgtgac ccatcagcgt tgcttaatta attgatgaca aattaaatgt
     1921 catcaatata atatgctctg caattattat acaaagcaat taaaacaagc ggataaaagg
     1981 acttgctttc aacccacccc taagtttaat agttactgag ggggatccac tagtgagctc
     2041 atgcatgatc tcgaattagc ttcaaaagcg ctctgaagtt cctatacttt ctagagaata
     2101 ggaacttcgg aataggaact tcaagatccc ctgattccct ttgtcaacag caatggataa
     2161 ttcgatttaa caaatgcatg gcgcaagggc tgctaaagga agcggaacac gtagaaagcc
     2221 agtccgcaga aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg
     2281 gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta
     2341 gactgggcgg ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt
     2401 aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg
     2461 cgcaggggat caagatctga tcaagagaca ggatgagggg tacccgcacg aacccagttg
     2521 acataagcct gttcggttcg taaactgtaa tgcaagtagc gtatgcgctc acgcaactgg
     2581 tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt
     2641 tatgactgtt tttttgtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc
     2701 gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct
     2761 aaaacaaagt taggcagccg ttgtgctggt gctttctgat agttgttgtg gggtaggcag
     2821 tcagagctcg atttgcttgt cgccataata gattcacaag aaggattcga catgggtcaa
     2881 agtag
//
