LOCUS       sequence-201165-        3440 bp    DNA     circular SYN 15-APR-2025
DEFINITION  LI genomic downstream module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3440)
  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 3440)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 Rsc203337 Downstream
FEATURES             Location/Qualifiers
     source          1..3440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1112..1128)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1112..1128)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1125..1147)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1136..1152
                     /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(1160..1166,1167..1184,1185..1190))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1205..1226
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1343..1360)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1514..2102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1594..1613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2217..2222,2223..2239,2240..2245)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2415..2948
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     3035..3053
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3091..3113)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3213..3232
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(3426..3440,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3440,1..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 ctcatctgtc gatgtcttgg ggccagcgat
      121 tgtatcgggg gatactcccc atcagctatt cagaaccctc aagacaactc tggacatcag
      181 cggctgccac ttgtccaaat cgcatcatgg caacgagaca ttggtggttg ctatggagag
      241 agggcttgca cgccaaaagc cagcgtcgtc cctcactcct caacctcgct gcgaaccatc
      301 agcacaggtt tgttcgtctt gcgcactacg ccttcggcca cactgccaag caccaggtgt
      361 ttgagcccgc gccggccgtg agtaccaagc acaatcaggt cagtgcccgt ctcgttggcc
      421 tcggccacaa tcgttgcgga aatctgcccc ggcacaatgg atttctctag caacttggtc
      481 gaatgcttga cgccagcggc ctccaatttg gtcgcaacct cggccagcgt gtctcggccg
      541 aactcgacaa cctgtttggc gtaggcaacg gagtcagccc cgaccatgtc gaaggtcacg
      601 tcgctgctat cggcaacaaa gagggcttcg atttcggcgc ccgtcgcttt gctgaccaca
      661 atcgcctggt gcagggcgag ataggacgcg tggctgccgt cgatggcgag caggatgcgt
      721 tggtacatga cggtttccaa aggcgttgtg cgggacagtg ccgcatgtta ccagcgtagg
      781 cggcaggatg ctgggctcat tgacccatat caacgctctt ccggggcgat tcccaatacc
      841 gccgcgcagg caatgcgccc ggcacgcagg ttgtccagag cggtgttggt ttcctgcagt
      901 cgatacggca tgacgtgaac ctgcaggggt actgcagccg ccacgtgcat gagcgccagg
      961 ccatccgatc gcgtcaggtt agccactgag acgagcttgc gctcctccca cagcagccgg
     1021 tacggaaacg tggggatgtc gctcatgggg ctgccaccgc aaaccacgga aggtgagacc
     1081 tctgcctgca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1141 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1201 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1261 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1321 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1381 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1441 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1501 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1561 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1621 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1681 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1741 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1801 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1861 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1921 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1981 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2041 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2101 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2161 aagggatttt ggtcatgagg cgcacaccgt ggaaacggat gaaggcacga acccagttga
     2221 cataagcctg ttcggttcgt aaactgtaat gcaagtagcg tatgcgctca cgcaactggt
     2281 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
     2341 atgactgttt ttttgtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
     2401 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta
     2461 aaacaaagtt aggtggctca agtatgggca tcattcgcac atgtaggctc ggccctgacc
     2521 aagtcaaatc catgcgggct gctcttgatc ttttcggtcg tgagttcggc gacgtagcca
     2581 cctactccca acatcagccg gactccgatt acctcgggaa cttgctccgt agtaagacat
     2641 tcatcgcgct tgctgccttc gaccaagaag cggttgttgg cgctctcgcg gcttacgttc
     2701 tgcccaagtt tgagcagccg cgtagtgaga tctatatcta tgatctcgca gtctccggag
     2761 agcaccggag gcagggcatt gccaccgcgc tcatcaatct cctcaagcat gaggccaacg
     2821 cgcttggtgc ttatgtgatc tacgtgcaag cagattacgg tgacgatccc gcagtggctc
     2881 tctatacaaa gttgggcata cgggaagaag tgatgcactt tgatatcgac ccaagtaccg
     2941 ccacctaaca attcgttcaa gccgagatcg gcttcccggc cgcggagttg ttcggtaaat
     3001 tgtcacaacg ccgctcatga cattaaccta taaaaatagg cgtatcacga ggcccttttg
     3061 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagaaggt
     3121 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     3181 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     3241 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     3301 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     3361 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     3421 gttttcccag tcacgacgtt
//
