LOCUS       sequence-201164-        3450 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LI genomic upstream module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3450)
  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 3450)
  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 Rsc202375 Upstream
FEATURES             Location/Qualifiers
     source          1..3450
                     /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(1103..1119)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1103..1119)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1116..1138)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1127..1143
                     /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(1151..1157,1158..1175,1176..1181))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1196..1217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1334..1351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1505..2093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1585..1604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2208..2213,2214..2230,2231..2236)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2425..2958
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     3045..3063
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3101..3123)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3223..3242
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(3436..3450,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3450,1..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 ctcacacctg accgagcaga tgccggtggt
      121 gacggtcgcg cccaacgacg cgctgctgga gaagctcaag tccaacatcc aggaagtgcg
      181 cgcgcgcggc ggcaagctct acgtgttcgc cgatgccgac acgcatatcc agtcgagcga
      241 cggcatccag gtgatccgca tgcccgagca ctatgggcag ctgtcgccga tcctgcacgt
      301 ggtgccgctg cagctgctgg cgtatcacac cgccgtggcg cgcgggacgg atgtggataa
      361 gccgcggaat ttggctaagt ctgtgacggt ggagtgaggg ggcggtagtg ctgttgtcct
      421 tgtaacggca gacaaaaaag tctgtaccag gtccgcagtt gaaacgtata ggcaccggtc
      481 tgccccgatt aagcgggggg ccggtgtttt tttcgaatag gtgttggctc agaggaggcg
      541 ggcgccgccg ctgtttccca gaccttagtc gtagggtgac cagcggaccc ggtcgtagca
      601 ggccgacggc aattcgatgc tcaacgttcc gaatttatcc agcagcttct tggcgagctt
      661 gggtgacgcg acctgttccg cgtataaggt gactaggtgt ctcacctcat atcggcgcac
      721 tttgcgatcg cttttccgat cgaccgactg agacgattca acgttttctt cgggaaagca
      781 cgctgtttcc gctatttgag atgttcactg gggcgcaatt ctctgcccgg gaagaacggg
      841 gccgcatcga tgcggtactt caaagcctcc caaagcgcat ggttggcgca ttcggggggc
      901 cgactcttgt gtccgcttga tgccacgtct cggatcgtga gccccggtcg atagttggag
      961 cctcttccac tgcccaggcc ggcggctcgc catcgttcta tatcggcaat catgatgttg
     1021 cggcgcttct tcataactga tcagctcaat cctgtcgccc tacgcgagac ctctgcctgc
     1081 atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1141 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1201 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1261 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1321 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1381 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1441 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1501 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1561 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1621 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1681 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1741 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1801 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1861 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1921 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1981 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2041 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2101 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2161 tggtcatgag gcgcacaccg tggaaacgga tgaaggcacg aacccagttg acataagcct
     2221 gttcggttcg taaactgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct
     2281 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt
     2341 tttttgtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat
     2401 gtttgatgtt atggagcagc aacgatgtta cgcagcagca acgatgttac gcagcagggc
     2461 agtcgcccta aaacaaagtt aggtggctca agtatgggca tcattcgcac atgtaggctc
     2521 ggccctgacc aagtcaaatc catgcgggct gctcttgatc ttttcggtcg tgagttcggc
     2581 gacgtagcca cctactccca acatcagccg gactccgatt acctcgggaa cttgctccgt
     2641 agtaagacat tcatcgcgct tgctgccttc gaccaagaag cggttgttgg cgctctcgcg
     2701 gcttacgttc tgcccaagtt tgagcagccg cgtagtgaga tctatatcta tgatctcgca
     2761 gtctccggag agcaccggag gcagggcatt gccaccgcgc tcatcaatct cctcaagcat
     2821 gaggccaacg cgcttggtgc ttatgtgatc tacgtgcaag cagattacgg tgacgatccc
     2881 gcagtggctc tctatacaaa gttgggcata cgggaagaag tgatgcactt tgatatcgac
     2941 ccaagtaccg ccacctaaca attcgttcaa gccgagatcg gcttcccggc cgcggagttg
     3001 ttcggtaaat tgtcacaacg ccgctcatga cattaaccta taaaaatagg cgtatcacga
     3061 ggcccttttg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     3121 cggagaaggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     3181 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     3241 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3301 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     3361 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     3421 taacgccagg gttttcccag tcacgacgtt
//
