LOCUS       sequence-201161-        3304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LI resistance gene module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3304)
  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 3304)
  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 F-G Kan
FEATURES             Location/Qualifiers
     source          1..3304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(99..914)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(99..914)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     822..841
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     complement(957..973)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(957..973)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(970..992)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    981..997
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1005..1011,1012..1029,1030..1035))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1050..1071
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1188..1205)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1359..1947)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1439..1458)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2062..2067,2068..2084,2085..2090)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2279..2812
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     2899..2917
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2955..2977)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3077..3096
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(3290..3304,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3304,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttcgagctcg gtacctcgcg aatgcatcta gatatcggat
       61 cccgggcccg tcgactgcag aggtacgggt ctcctgagtt agaaaaactc atcgagcatc
      121 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
      181 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
      241 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
      301 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
      361 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
      421 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
      481 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
      541 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
      601 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
      661 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
      721 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
      781 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
      841 ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt
      901 tgaatatggc tcattgtcag agacctctgc ctgcatgcaa gcttggcgta atcatggtca
      961 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     1021 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     1081 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     1141 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     1201 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1261 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1321 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1381 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1441 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1501 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1561 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1621 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1681 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1741 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     1801 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1861 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1921 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1981 gctcagtgga acgaaaactc acgttaaggg attttggtca tgaggcgcac accgtggaaa
     2041 cggatgaagg cacgaaccca gttgacataa gcctgttcgg ttcgtaaact gtaatgcaag
     2101 tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc
     2161 gcagtggcgg ttttcatggc ttgttatgac tgtttttttg tacagtctat gcctcgggca
     2221 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat
     2281 gttacgcagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaggtgg
     2341 ctcaagtatg ggcatcattc gcacatgtag gctcggccct gaccaagtca aatccatgcg
     2401 ggctgctctt gatcttttcg gtcgtgagtt cggcgacgta gccacctact cccaacatca
     2461 gccggactcc gattacctcg ggaacttgct ccgtagtaag acattcatcg cgcttgctgc
     2521 cttcgaccaa gaagcggttg ttggcgctct cgcggcttac gttctgccca agtttgagca
     2581 gccgcgtagt gagatctata tctatgatct cgcagtctcc ggagagcacc ggaggcaggg
     2641 cattgccacc gcgctcatca atctcctcaa gcatgaggcc aacgcgcttg gtgcttatgt
     2701 gatctacgtg caagcagatt acggtgacga tcccgcagtg gctctctata caaagttggg
     2761 catacgggaa gaagtgatgc actttgatat cgacccaagt accgccacct aacaattcgt
     2821 tcaagccgag atcggcttcc cggccgcgga gttgttcggt aaattgtcac aacgccgctc
     2881 atgacattaa cctataaaaa taggcgtatc acgaggccct tttgtctcgc gcgtttcggt
     2941 gatgacggtg aaaacctctg acacatgcag ctcccggaga aggtcacagc ttgtctgtaa
     3001 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     3061 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     3121 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     3181 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     3241 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     3301 cgtt
//
