LOCUS       sequence-443284-        5075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pDonorTp with addition of fugfp gene from pUS252 outside of the FRT 
            sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5075)
  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 5075)
  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-fugfp
FEATURES             Location/Qualifiers
     source          1..5075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(118..151)
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     protein_bind    178..225
                     /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)."
     CDS             complement(397..633)
                     /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"
     promoter        complement(join(959..964,965..981,982..987))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    1383..1430
                     /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)."
     oriT            1873..1981
                     /note="incP origin of transfer"
     rep_origin      2032..2388
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     terminator      2732..2826
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      2929..3015
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             3976..3987
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 gaaacggtct aaataggctg attcaaggca tttacgggag aaaaaatcgg ctcaaacatg
       61 aagaaatgaa atgactgagt cagccgagaa gaatttcccc gcttattcgc accttcccta
      121 gtattatacc taggactgag ctagctgtca acccggggat ctcgaattgg ggatcttgaa
      181 gtacctattc cgaagttcct attctctaga aagtatagga acttcagagc gcttttgaag
      241 ctgatgtgct taaaaactta ctcaatggaa taattctaga taattcacta gtgattggaa
      301 atgttggatc ctcatactct tccctcgagt agaattctga ctcttataca caagtgcggc
      361 cgcactagtc tagatttaaa ttaattaaga gctcgcttag gccacacgtt caagtgcagc
      421 cacaggataa atttgcactg agcctgggtg ggattcggac tcgaccgcat agccttcagg
      481 agtgagtttt gtgcaatacc aaccgacgac ttgaccctgc caagcggcac cagatttctt
      541 gcgtacgcga tcccctaagc caaaggtggc actcagggga agcgcaaact gccctgcaac
      601 gggagcgttg gcttcatcgc tactttgacc catgtcgaat ccttcttgtg aatctattat
      661 ggcgacaagc aaatcgagct ctgactgcct accccacaac aactatcaga aagcaccagc
      721 acaacggctg cctaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct
      781 ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat
      841 agactgtaca aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc
      901 gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag
      961 tttacgaacc gaacaggctt atgtcaactg ggttcgtgcg ggtacccctc atcctgtctc
     1021 ttgatcagat cttgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt
     1081 tactttgcag ggcttcccaa ccttaccaga gggcgcccca gctggcaatt ccggttcgct
     1141 tgctgtccat aaaaccgccc agtctagcta tcgccatgta agcccactgc aagctacctg
     1201 ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg
     1261 gggtcagcac cgtttctgcg gactggcttt ctacgtgttc cgcttccttt agcagccctt
     1321 gcgccatgca tttgttaaat cgaattatcc attgctgttg acaaagggaa tcaggggatc
     1381 ttgaagttcc tattccgaag ttcctattct ctagaaagta taggaacttc agagcgcttt
     1441 tgaagctaat tcgagatcat gcatgagctc actagtggat ccccctcagt aactattaaa
     1501 cttaggggtg ggttgaaagc aagtcctttt atccgcttgt tttaattgct ttgtataata
     1561 attgcagagc atattatatt gatgacattt aatttgtcat caattaatta agcaacgctg
     1621 atgggtcacg acgacaatta aatagtcaca atgacattaa tctgtcaccg acgacagata
     1681 atttgtcact gtacactacg ccttttgtgg agatgtctaa taaactaaaa attttttact
     1741 ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt
     1801 tttcgcacga tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca
     1861 acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt
     1921 cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc
     1981 gataagctct gataacgcag gaaagaacat gggcagttca acctgttgat agtacgtact
     2041 aagctctcat gtttcacgta ctaagctctc aagtttaacg tactaagctc tcatgtttaa
     2101 cgaactaaac cctcatggct aacgtactaa gctctcatgg ctaacgtact aagctctcat
     2161 gtttcacgta ctaagctctc atgtttgaac aataaaatta atataaatca gcaacttaaa
     2221 tagcctctaa ggttttaagt tttataagaa aaaaaagaat atataaggct tttaaagctt
     2281 ttaaggttta acggttgtgg acaacaagcc agggatgtaa cgcactgaga agcccttaga
     2341 gcctctcaaa gcaattttga gtgacacagg aacacttaac ggctgacaac cggctggtaa
     2401 caggattagc agagcgaggt atgtaggcgg tgctacaaga ttgtactgag agtgcaccat
     2461 acaaaaccat tgtagagagg tagcattgct acgtctctac gtgtacagtg actaattata
     2521 tgtcgttgtg acaaattatt gtcatcagta aaatccttat acagtataga ttatagcgct
     2581 ttggcagttt tagcataacc tctttgcagt gacaaaatag atgtcgttgt ccgtgattgt
     2641 gacaaattag ctgtcgcttt gcaagatagg aaaaagcttt tgtgtatttt cataatgaca
     2701 aattgactgt cgccttaatt agctgagctt ggactcctgt tgatagatcc agtaatgacc
     2761 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
     2821 gagaatccaa gctagactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt
     2881 attggtgccc ttaaacgcct ggggtaatga ctctctagct tgaggcatca aataaaacga
     2941 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     3001 ctgagtagga caaatccgcc gctaggcata tgtaattcgc tgttaaaagg acaattacaa
     3061 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     3121 gaatcaggat attcttctaa tacctggaat gctgttttgc cggggaatta catatgggaa
     3181 acagctatga cggtaccaaa aaaaaccccg ccctgtcagg ggcggggttt ttttttgagc
     3241 tcttattagc tcttatacag ctcgtccata cccaggctca agcccgcagc ggtcacaaat
     3301 tccaccagga ccatgtgatc acggcgttcc tgcggatctt tgctcagcac gctattggtc
     3361 gtcagatagt gttgttccgg cagcaacacc ggaccctcac caattggggt gttctgttgg
     3421 taatggtcgg ccagttggac gctaccgtca tcaacattgt ggcgaatacg gaattgggcc
     3481 tccaagccgt tacgattctt gtctgccgta atgtaaacat aatggctgtt gaagctatac
     3541 tccagtttat ggcccagaat gttaccatcc tctttaaact ccaaaccctt caaatcgata
     3601 cgattgacca gcgcttcacc ttcaaactta acctctgcgc gagttttgta cgtgccatct
     3661 tccttgaagg aaatggtacg ctcctggacg taaccgtccg gcatggcact tttaaagaaa
     3721 tcgttttggc gcatgtgctc cggatacttc gcaaaacact gcacgccata cgacaaggtc
     3781 gtcaccaagg tcggccaagg caccggcagg cgaccggtcg tgcagatgaa cttaatttcc
     3841 agtttgccgt tgctcgcgtc gccataacct tcaccgcgaa cgctaaaacg atgaccgttc
     3901 acgtcgccct ccagctcgat cagaatcgga accaagcccg agaaaatatc ttcgccactg
     3961 ctcaccatgg gactctttct cctctttaat ctactaggga aggtgcgaac aagtccctga
     4021 tatgagatca tgtttgtcat ctggagccat agaacagggt tcatcatgag tcatcaactt
     4081 accttcgccg acagtgaatt cagcagtaag cgccgtcaga ccagaaaaga gattttcttg
     4141 tcccgcatgg agcagattct gccatggcaa aacatggtgg aagtcatcga gccgttttac
     4201 cccaaggctg gtaatggccg gcgaccttat ccgctggaaa ccatgctacg cattcactgc
     4261 atgcagcatt ggtacaacct gagcgatggc gcgatggaag atgctctgta cgaaatcgcc
     4321 tccatgcgtc tgtttgcccg gttatccctg gatagcgcct tgccggaccg caccaccatc
     4381 atgaatttcc gccacctgct ggagcagcat caactggccc gccaattgtt caagaccatc
     4441 aatcgctggc tggccgaagc aggcgtcatg atgactcaag gcaccttggt cgatgccacc
     4501 atcattgagg cacccagctc gaccaagaac aaagagcagc aacgcgatcc ggagatgcat
     4561 cagaccaaga aaggcaatca gtggcacttt ggcatgaagg cccacattgg tgtcgatgcc
     4621 aagagtggcc tgacccacag cctggtcacc accgcggcca acgagcatga cctcaatcag
     4681 ctgggtaatc tgctgcatgg agaggagcaa tttgtctcag ccgatgccgg ctaccaaggg
     4741 gcgccacagc gcgaggagct ggccgaggtg gatgtggact ggctgatcgc cgagcgcccc
     4801 ggcaaggtaa gaaccttgaa acagcatcca cgcaagaaca aaacggccat caacatcgaa
     4861 tacatgaaag ccagcatccg ggccagggtg gagcacccat ttcgcatcat caagcgacag
     4921 ttcggcttcg tgaaagccag atacaagggg ttgctgaaaa acgataacca actggcgatg
     4981 ttattcacgc tggccaacct gtttcgggcg gaccaaatga tacgtcagtg ggagagatct
     5041 cactaaaaac tggggataac gccttaaatg gcgaa
//
