LOCUS       sequence-201160-        4021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LI resistance gene module. Please note that the plasmid contains 
            Apramycin selection, not Gentamycin..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4021)
  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 4021)
  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 Gent
FEATURES             Location/Qualifiers
     source          1..4021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             174..974
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     promoter        complement(join(1549..1555,1556..1573,1574..1579))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1594..1615
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1735..1752)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1906..2494)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1986..2005)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2665..3456,3457..3525))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3288..3307
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3526..3630)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3698..3716
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 taatacgact cactataggg agagcggccg ccagatcttc cggatggctc gagtttttca
       61 gcaagatttg gtctcctgag ggttcatgta cagctccatc agcaaaaggg gatgataagt
      121 ttatcaccac cgactatttg caacagtgcc gttgatcgtg ctatgatcga ctgatgtcat
      181 cagcggtgga gtgcaatgtc gtgcaatacg aatggcgaaa agccgagctc atcggtcagc
      241 ttctcaacct tggggttacc cccggcggtg tgctgctggt ccacagctcc ttccgtagcg
      301 tccggcccct cgaagatggg ccacttggac tgatcgaggc cctgcgtgct gcgctgggtc
      361 cgggagggac gctcgtcatg ccctcgtggt caggtctgga cgacgagccg ttcgatcctg
      421 ccacgtcgcc cgttacaccg gaccttggag ttgtctctga cacattctgg cgcctgccaa
      481 atgtaaagcg cagcgcccat ccatttgcct ttgcggcagc ggggccacag gcagagcaga
      541 tcatctctga tccattgccc ctgccacctc actcgcctgc aagcccggtc gcccgtgtcc
      601 atgaactcga tgggcaggta cttctccttg gcgtgggaca cgatgccaac acgacgctgc
      661 atcttgccga gttgatggca aaggttccct atggggtgcc gagacactgc accattcttc
      721 aggatggcaa gttggtacgc gtcgattatc tcgagaatga ccactgctgt gagcgctttg
      781 ccttggcgga caggtggctc aaggagaaga gccttcagaa ggaaggtcca gtcggtcatg
      841 cctttgctcg gttgatccgc tcccgcgaca ttgtggcgac agccctgggt caactgggcc
      901 gagatccgtt gatcttcctg catccgccag aggcgggatg cgaagaatgc gatgccgctc
      961 gccagtcgat tggctgagct catgagcgga gaacgagatg acgttggagg ggcaaggtcg
     1021 cgctgattgc tggggcaaca cgtggagcgg atcggggatt gtctttcttc agctcgctga
     1081 tgatatgctg acgctcaatg cctgtcagag accaatcttt ctagaagatc tcctacaata
     1141 ttctcagctg ccatggaaaa tcgatgttct tcttttattc tctcaagatt ttcaggctgt
     1201 atattaaaac ttatattaag aactatgcta accacctcat caggaaccgt tgtaggtggc
     1261 gtgggttttc ttggcaatcg actctcatga aaactacgag ctaaatattc aatatgttcc
     1321 tcttgaccaa ctttattctg catttttttt gaacgaggtt tagagcaagc ttcaggaaac
     1381 tgagacagga attttattaa aaatttaaat tttgaagaaa gttcagggtt aatagcatcc
     1441 attttttgct ttgcaagttc ctcagcattc ttaacaaaag acgtctcttt tgacatgttt
     1501 aaagtttaaa cctcctgtgt gaaattatta tccgctcata attccacaca ttatacgagc
     1561 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1621 gttgcgctca ctgccaattg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     1681 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1741 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1801 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1861 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1921 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1981 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2041 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2101 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2161 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2221 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2281 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2341 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2401 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2461 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2521 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2581 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2641 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2701 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     2761 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     2821 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     2881 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     2941 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3001 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3061 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3121 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3181 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3241 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3301 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3361 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3421 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3481 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     3541 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3601 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     3661 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccgcc
     3721 cctgcagccg aattatatta tttttgccaa ataattttta acaaaagctc tgaagtcttc
     3781 ttcatttaaa ttcttagatg atacttcatc tggaaaattg tcccaattag tagcatcacg
     3841 ctgtgagtaa gttctaaacc atttttttat tgttgtatta tctctaatct tactactcga
     3901 tgagttttcg gtattatctc tatttttaac ttggagcagg ttccattcat tgtttttttc
     3961 atcatagtga ataaaatcaa ctgctttaac acttgtgcct gaacaccata tccatccggc
     4021 g
//
