LOCUS       sequence-198155-        1718 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  LI Gateway cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1718)
  AUTHORS   Wu D, Schandry N, Lahaye T
  TITLE     A modular toolbox for Golden-Gate-based plasmid assembly streamlines
            generation of Ralstonia solanacearum species complex knockout 
            strains and multi-cassette complementation constructs.
  JOURNAL   Mol Plant Pathol. 2017 Oct 27. doi: 10.1111/mpp.12632.
  PUBMED    29077245
REFERENCE   2  (bases 1 to 1718)
  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: "Mol Plant 
            Pathol. 2017 Oct 27. doi: 10.1111/mpp.12632."
COMMENT     Sequence Label: LI C-D Dest
FEATURES             Location/Qualifiers
     source          1..1718
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    1..125
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(162..167,168..185,186..192)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             246..905
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(312..331)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1247..1552
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1431..1450
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1593..1717)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg atataaatat caatatatta
       61 aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca tatccagtca
      121 ctatggcggc ccctcgaggg gccgcattag gcaccccagg ctttacactt tatgcttccg
      181 gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga gctaaggaag
      241 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
      301 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
      361 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
      421 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag
      481 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
      541 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
      601 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
      661 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
      721 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
      781 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
      841 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
      901 cgtaaagatc tggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg
      961 atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggta
     1021 tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc
     1081 atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt
     1141 ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt
     1201 attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc agtttaaggt
     1261 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     1321 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     1381 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     1441 caccgatatg gccagtgtgc cggtgtccgt tatcggggaa gaagtggctg atctcagcca
     1501 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     1561 ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt tgtgttttac
     1621 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     1681 tttacgtttc tcgttcagct ttcttgtaca aagtggtg
//
