LOCUS       sequence-198162-        4023 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 4023)
  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 4023)
  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..4023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     305..324
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        305..323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(join(1855..1861,1862..1879,1880..1885))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1900..1921
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2041..2058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2212..2800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2292..2311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2971..3762,3763..3831))
                     /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     3594..3613
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3832..3936)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4004..4022
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc
       61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc
      121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact
      181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt
      241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc
      301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt
      361 ttcagcaaga tttggtctcc tgagggttca tgtgcagctc catcagcaaa aggggatgat
      421 aagtttatca ccaccgacta tttgcaacag tgccgttgat cgtgctatga tcgactgatg
      481 tcatcagcgg tggagtgcaa tgtcgtgcaa tacgaatggc gaaaagccga gctcatcggt
      541 cagcttctca accttggggt tacccccggc ggtgtgctgc tggtccacag ctccttccgt
      601 agcgtccggc ccctcgaaga tgggccactt ggactgatcg aggccctgcg tgctgcgctg
      661 ggtccgggag ggacgctcgt catgccctcg tggtcaggtc tggacgacga gccgttcgat
      721 cctgccacgt cgcccgttac accggacctt ggagttgtct ctgacacatt ctggcgcctg
      781 ccaaatgtaa agcgcagcgc ccatccattt gcctttgcgg cagcggggcc acaggcagag
      841 cagatcatct ctgatccatt gcccctgcca cctcactcgc ctgcaagccc ggtcgcccgt
      901 gtccatgaac tcgatgggca ggtacttctc cttggcgtgg gacacgatgc caacacgacg
      961 ctgcatcttg ccgagttgat ggcaaaggtt ccctatgggg tgccgagaca ctgcaccatt
     1021 cttcaggatg gcaagttggt acgcgtcgat tatctcgaga atgaccactg ctgtgagcgc
     1081 tttgccttgg cggacaggtg gctcaaggag aagagccttc agaaggaagg tccagtcggt
     1141 catgcctttg ctcggttgat ccgctcccgc gacattgtgg cgacagccct gggtcaactg
     1201 ggccgagatc cgttgatctt cctgcatccg ccagagggcg ggatgcgaag aatgcgatgc
     1261 cgctcgccag tcgattggct gagctcatga gcggagaacg agatgacgtt ggaggggcaa
     1321 ggtcgcgctg attgctgggg caacacgtgg agcggatcgg ggattgtctt tcttcagctc
     1381 gctgatgata tgctgacgct caatgcctgt cagagaccaa atctttctag aagatctcct
     1441 acaatattct cagctgccat ggaaaatcga tgttcttctt ttattctctc aagattttca
     1501 ggctgtatat taaaacttat attaagaact atgctaacca cctcatcagg aaccgttgta
     1561 ggtggcgtgg gttttcttgg caatcgactc tcatgaaaac tacgagctaa atattcaata
     1621 tgttcctctt gaccaacttt attctgcatt ttttttgaac gaggtttaga gcaagcttca
     1681 ggaaactgag acaggaattt tattaaaaat ttaaattttg aagaaagttc agggttaata
     1741 gcatccattt tttgctttgc aagttcctca gcattcttaa caaaagacgt ctcttttgac
     1801 atgtttaaag tttaaacctc ctgtgtgaaa ttattatccg ctcataattc cacacattat
     1861 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     1921 aattgcgttg cgctcactgc caattgcttt ccagtcggga aacctgtcgt gccagctgca
     1981 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2041 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2101 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2161 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2221 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2281 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2341 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2401 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2461 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2521 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2581 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2641 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2701 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2761 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2821 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2881 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2941 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3001 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3061 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3121 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3181 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3241 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3301 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3361 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3421 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3481 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3541 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3601 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3661 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3721 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3781 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3841 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3901 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3961 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     4021 gcc
//
