LOCUS       sequence-201159-        3731 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 3731)
  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 3731)
  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 Spec
FEATURES             Location/Qualifiers
     source          1..3731
                     /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             355..1146
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     primer_bind     complement(1384..1400)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1384..1400)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1397..1419)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1408..1424
                     /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(1432..1438,1439..1456,1457..1462))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1477..1498
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1615..1632)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1786..2374)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1866..1885)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2489..2494,2495..2511,2512..2517)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2706..3239
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     3326..3344
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3382..3404)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3504..3523
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(3717..3731,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3731,1..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 tcctgagggc acgaacccag tggacataag cctgttcggt tcgtaagctg
      121 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
      181 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct
      241 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
      301 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa catcatgggg
      361 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
      421 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
      481 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
      541 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
      601 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
      661 gctaagcgcg aactacaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
      721 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
      781 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
      841 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
      901 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
      961 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
     1021 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc
     1081 gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc
     1141 aaataatgtc tagctagaaa ttcgttcaag ccgacgccgc ttcgcggcgc ggcttaactc
     1201 aagcgttaga tgcactaagc acataattgc tcacagccaa actatcaggt caagtctgct
     1261 tttattattt ttaagcgtgc ataataagcc ctacacaaat tgggagatat atcatgcatg
     1321 actgtcagag accagggggc ccgtcgactg cagaggcctg catgcaagct tggcgtaatc
     1381 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     1441 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     1501 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     1561 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1621 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1681 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1741 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1801 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1861 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1921 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1981 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2041 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2101 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2161 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2221 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2281 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2341 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2401 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga ggcgcacacc
     2461 gtggaaacgg atgaaggcac gaacccagtt gacataagcc tgttcggttc gtaaactgta
     2521 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
     2581 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgtac agtctatgcc
     2641 tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag
     2701 caacgatgtt acgcagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt
     2761 taggtggctc aagtatgggc atcattcgca catgtaggct cggccctgac caagtcaaat
     2821 ccatgcgggc tgctcttgat cttttcggtc gtgagttcgg cgacgtagcc acctactccc
     2881 aacatcagcc ggactccgat tacctcggga acttgctccg tagtaagaca ttcatcgcgc
     2941 ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc ggcttacgtt ctgcccaagt
     3001 ttgagcagcc gcgtagtgag atctatatct atgatctcgc agtctccgga gagcaccgga
     3061 ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca tgaggccaac gcgcttggtg
     3121 cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc cgcagtggct ctctatacaa
     3181 agttgggcat acgggaagaa gtgatgcact ttgatatcga cccaagtacc gccacctaac
     3241 aattcgttca agccgagatc ggcttcccgg ccgcggagtt gttcggtaaa ttgtcacaac
     3301 gccgctcatg acattaacct ataaaaatag gcgtatcacg aggccctttt gtctcgcgcg
     3361 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagaagg tcacagcttg
     3421 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     3481 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     3541 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     3601 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     3661 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     3721 gtcacgacgt t
//
