LOCUS       sequence-201163-        3229 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 3229)
  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 3229)
  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 Cml
FEATURES             Location/Qualifiers
     source          1..3229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(78..83,84..101,102..108)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             162..821
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(228..247)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(882..898)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(882..898)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(895..917)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    906..922
                     /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(930..936,937..954,955..960))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    975..996
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1113..1130)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1284..1872)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1364..1383)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1987..1992,1993..2009,2010..2015)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2204..2737
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     2824..2842
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2880..2902)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3002..3021
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(3215..3229,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3229,1..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 cccaatggtc tcatgagttt acactttatg cttccggctc gtataatgtg tggattttga
      121 gttaggatcc gtcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
      181 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
      241 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
      301 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
      361 tgaatgctca tccggaattc cgtatggcaa tgaaagacgg tgagctggtg atatgggata
      421 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
      481 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
      541 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag
      601 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
      661 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc
      721 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg
      781 aattacaaca gtactgcgat gagtggcagg gcggggcgta atgtcagaga ccaagggccc
      841 gtcgactgca gaggcctgca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg
      901 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
      961 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     1021 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1081 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1141 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1201 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1261 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1321 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1381 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1441 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1501 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1561 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1621 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1681 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1741 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1801 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1861 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1921 aactcacgtt aagggatttt ggtcatgagg cgcacaccgt ggaaacggat gaaggcacga
     1981 acccagttga cataagcctg ttcggttcgt aaactgtaat gcaagtagcg tatgcgctca
     2041 cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc
     2101 atggcttgtt atgactgttt ttttgtacag tctatgcctc gggcatccaa gcagcaagcg
     2161 cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagcaa
     2221 cgatgttacg cagcagggca gtcgccctaa aacaaagtta ggtggctcaa gtatgggcat
     2281 cattcgcaca tgtaggctcg gccctgacca agtcaaatcc atgcgggctg ctcttgatct
     2341 tttcggtcgt gagttcggcg acgtagccac ctactcccaa catcagccgg actccgatta
     2401 cctcgggaac ttgctccgta gtaagacatt catcgcgctt gctgccttcg accaagaagc
     2461 ggttgttggc gctctcgcgg cttacgttct gcccaagttt gagcagccgc gtagtgagat
     2521 ctatatctat gatctcgcag tctccggaga gcaccggagg cagggcattg ccaccgcgct
     2581 catcaatctc ctcaagcatg aggccaacgc gcttggtgct tatgtgatct acgtgcaagc
     2641 agattacggt gacgatcccg cagtggctct ctatacaaag ttgggcatac gggaagaagt
     2701 gatgcacttt gatatcgacc caagtaccgc cacctaacaa ttcgttcaag ccgagatcgg
     2761 cttcccggcc gcggagttgt tcggtaaatt gtcacaacgc cgctcatgac attaacctat
     2821 aaaaataggc gtatcacgag gcccttttgt ctcgcgcgtt tcggtgatga cggtgaaaac
     2881 ctctgacaca tgcagctccc ggagaaggtc acagcttgtc tgtaagcgga tgccgggagc
     2941 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     3001 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     3061 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     3121 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     3181 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgtt
//
