LOCUS       sequence-201156-        4273 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LI Gene of Interest module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4273)
  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 4273)
  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 GUS
FEATURES             Location/Qualifiers
     source          1..4273
                     /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             77..1885
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     primer_bind     complement(1945..1961)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1945..1961)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1958..1980)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1969..1985
                     /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(1993..1999,2000..2017,2018..2023))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2038..2059
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2176..2193)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2347..2935)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2427..2446)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(3050..3055,3056..3072,3073..3078)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3248..3781
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     3868..3886
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3924..3946)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4046..4065
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4259..4273,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4273,1..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 cccttggtct cgcaccatgt tacgtcctgt agaaacccca acccgtgaaa tcaaaaaact
      121 cgacggcctg tgggcattca gtctggatcg cgaaaactgt ggaattgatc agcgttggtg
      181 ggaaagcgcg ttacaagaaa gccgggcaat tgctgtgcca ggcagtttta acgatcagtt
      241 cgccgatgca gatattcgta attatgcggg caacgtctgg tatcagcgcg aagtctttat
      301 accgaaaggt tgggcaggcc agcgtatcgt gctgcgtttc gatgcggtca ctcattacgg
      361 caaagtgtgg gtcaataatc aggaagtgat ggagcatcag ggcggctata cgccatttga
      421 agccgatgtc acgccgtatg ttattgccgg gaaaagtgta cgtatcaccg tttgtgtgaa
      481 caacgaactg aactggcaga ctatcccgcc gggaatggtg attaccgacg aaaacggcaa
      541 gaaaaagcag tcttacttcc atgatttctt taactatgcc ggaatccatc gcagcgtaat
      601 gctctacacc acgccgaaca cctgggtgga cgatatcacc gtggtgacgc atgtcgcgca
      661 agactgtaac cacgcgtctg ttgactggca ggtggtggcc aatggtgatg tcagcgttga
      721 actgcgtgat gcggatcaac aggtggttgc aactggacaa ggcactagcg ggactttgca
      781 agtggtgaat ccgcacctct ggcaaccggg tgaaggttat ctctatgaac tgtgcgtcac
      841 agccaaaagc cagacagagt gtgatatcta cccgcttcgc gtcggcatcc ggtcagtggc
      901 agtgaagggc gaacagttcc tgattaacca caaaccgttc tactttactg gctttggtcg
      961 tcatgaagat gcggacttgc gtggcaaagg attcgataac gtgctgatgg tgcacgacca
     1021 cgcattaatg gactggattg gggccaactc ctaccgtacc tcgcattacc cttacgctga
     1081 agagatgctc gactgggcag atgaacatgg catcgtggtg attgatgaaa ctgctgctgt
     1141 cggctttaac ctctctttag gcattggttt cgaagcgggc aacaagccga aagaactgta
     1201 cagcgaagag gcagtcaacg gggaaactca gcaagcgcac ttacaggcga ttaaagagct
     1261 gatagcgcgt gacaaaaacc acccaagcgt ggtgatgtgg agtattgcca acgaaccgga
     1321 tacccgtccg caaggtgcac gggaatattt cgcgccactg gcggaagcaa cgcgtaaact
     1381 cgacccgacg cgtccgatca cctgcgtcaa tgtaatgttc tgcgacgctc acaccgatac
     1441 catcagcgat ctctttgatg tgctgtgcct gaaccgttat tacggatggt atgtccaaag
     1501 cggcgatttg gaaacggcag agaaggtact ggaaaaagaa cttctggcct ggcaggagaa
     1561 actgcatcag ccgattatca tcaccgaata cggcgtggat acgttagccg ggctgcactc
     1621 aatgtacacc gacatgtgga gtgaagagta tcagtgtgca tggctggata tgtatcaccg
     1681 cgtctttgat cgcgtcagcg ccgtcgtcgg tgaacaggta tggaatttcg ccgattttgc
     1741 gacctcgcaa ggcatattgc gcgttggcgg taacaagaaa gggatcttca ctcgcgaccg
     1801 caaaccgaag tcggcggctt ttctgctgca aaaacgctgg actggcatga acttcggtga
     1861 aaaaccgcag cagggaggca aacaaaaggc gagaccaggg cccgtcgact gcagaggcct
     1921 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     1981 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2041 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2101 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2161 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2221 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2281 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2341 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2401 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2461 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2521 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2581 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2641 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2701 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2761 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2821 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2881 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2941 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3001 tttggtcatg aggcgcacac cgtggaaacg gatgaaggca cgaacccagt tgacataagc
     3061 ctgttcggtt cgtaaactgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac
     3121 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
     3181 tttttttgta cagtctatgc ctcgggcatc caagcagcaa gcgskttrmk sykwkggwss
     3241 akswwygatg ttacgcagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa
     3301 gttaggtggc tcaagtatgg gcatcattcg cacatgtagg ctcggccctg accaagtcaa
     3361 atccatgcgg gctgctcttg atcttttcgg tcgtgagttc ggcgacgtag ccacctactc
     3421 ccaacatcag ccggactccg attacctcgg gaacttgctc cgtagtaaga cattcatcgc
     3481 gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc gcggcttacg ttctgcccaa
     3541 gtttgagcag ccgcgtagtg agatctatat ctatgatctc gcagtctccg gagagcaccg
     3601 gaggcagggc attgccaccg cgctcatcaa tctcctcaag catgaggcca acgcgcttgg
     3661 tgcttatgtg atctacgtgc aagcagatta cggtgacgat cccgcagtgg ctctctatac
     3721 aaagttgggc atacgggaag aagtgatgca ctttgatatc gacccaagta ccgccaccta
     3781 acaattcgtt caagccgaga tcggcttccc ggccgcggag ttgttcggta aattgtcaca
     3841 acgccgctca tgacattaac ctataaaaat aggcgtatca cgaggccctt ttgtctcgcg
     3901 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagaa ggtcacagct
     3961 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     4021 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     4081 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
     4141 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     4201 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     4261 cagtcacgac gtt
//
