LOCUS       sequence-201170-        2673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LI terminator module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2673)
  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 2673)
  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 E-F Tfd
FEATURES             Location/Qualifiers
     source          1..2673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      77..123
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     primer_bind     complement(326..342)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(326..342)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(339..361)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    350..366
                     /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(374..380,381..398,399..404))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    419..440
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(557..574)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(728..1316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(808..827)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1431..1436,1437..1453,1454..1459)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1648..2181
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     2268..2286
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2324..2346)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2446..2465
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(2659..2673,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(2673,1..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 caaatcccga tacaattaaa ggctcctttt ggagcctttt ttttggagat
      121 tttcaacgtg aaaaaattat tattcgcaat tcctttagtt gttcctttct attctcactc
      181 cgctgaaact gttgaaagtt gtttagcaaa acctcataca gaaaattcat ttactaacgt
      241 ctggaaagac gacaaaactt tagagtgaga gagaccaagg gcccgtcgac tgcagaggcc
      301 tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
      361 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
      421 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
      481 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
      541 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      601 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
      661 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      721 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      781 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      841 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      901 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      961 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1021 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1081 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1141 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1201 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1261 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1321 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1381 ttttggtcat gaggcgcaca ccgtggaaac ggatgaaggc acgaacccag ttgacataag
     1441 cctgttcggt tcgtaaactg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa
     1501 ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact
     1561 gtttttttgt acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc
     1621 gatgtttgat gttatggagc agcaacgatg ttacgcagca gcaacgatgt tacgcagcag
     1681 ggcagtcgcc ctaaaacaaa gttaggtggc tcaagtatgg gcatcattcg cacatgtagg
     1741 ctcggccctg accaagtcaa atccatgcgg gctgctcttg atcttttcgg tcgtgagttc
     1801 ggcgacgtag ccacctactc ccaacatcag ccggactccg attacctcgg gaacttgctc
     1861 cgtagtaaga cattcatcgc gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc
     1921 gcggcttacg ttctgcccaa gtttgagcag ccgcgtagtg agatctatat ctatgatctc
     1981 gcagtctccg gagagcaccg gaggcagggc attgccaccg cgctcatcaa tctcctcaag
     2041 catgaggcca acgcgcttgg tgcttatgtg atctacgtgc aagcagatta cggtgacgat
     2101 cccgcagtgg ctctctatac aaagttgggc atacgggaag aagtgatgca ctttgatatc
     2161 gacccaagta ccgccaccta acaattcgtt caagccgaga tcggcttccc ggccgcggag
     2221 ttgttcggta aattgtcaca acgccgctca tgacattaac ctataaaaat aggcgtatca
     2281 cgaggccctt ttgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     2341 tcccggagaa ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     2401 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     2461 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     2521 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     2581 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     2641 gggtaacgcc agggttttcc cagtcacgac gtt
//
