LOCUS       sequence-201162-        4442 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 4442)
  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 4442)
  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 Tet
FEATURES             Location/Qualifiers
     source          1..4442
                     /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             complement(78..1277)
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     CDS             1383..2033
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     primer_bind     complement(2095..2111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2095..2111)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2108..2130)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2119..2135
                     /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(2143..2149,2150..2167,2168..2173))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2188..2209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2326..2343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2497..3085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2577..2596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(3200..3205,3206..3222,3223..3228)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3417..3950
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     4037..4055
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4093..4115)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4215..4234
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4428..4442,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4442,1..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 cccattggtc tctgacatca gcgatcggct cgttgccctg cgccgctcca aagcccgcga
      121 cgcagcgccg gcaggcagag caagtagagg gcagcgcctg caatccatgc ccacccgttc
      181 cacgttgtta tagaagccgc atagatcgcc gtgaagagga ggggtccgac gatcgaggtc
      241 aggctggtga gcgccgccag tgagccttgc agctgcccct gacgttcctc atccacctgc
      301 ctggacaaca ttgcttgcag cgccggcatt ccgatgccac ccgaagcaag caggaccatg
      361 atcgggaacg ccatccatcc ccgtgtcgcg aaggcaagca ggatgtagcc tgtgccgtcg
      421 gcaatcattc cgagcatgag tgcccgcctt tcgccgagcc gggcggctac agggccggtg
      481 atcattgcct gggcgagtga atgcagaatg ccaaatgcgg caagcgaaat gccgatcgtg
      541 gtcgcgtccc agtgaaagcg atcctcgccg aaaatgaccc aaagcgcggc cggcacctgt
      601 ccgacaagtt gcatgatgaa gaagaccgcc atcagggcgg cgacgacggt catgccccgg
      661 gcccaccgga acgaagcgag cgggttgaga gcctcccggc gtaacggccg gcgttcgcct
      721 ttgtgcgact ccggcaaaag gaaacagccc gtcaggaaat tgaggccgtt caaggctgcc
      781 gcggcgaaga acggagcgtg gggggagaaa ccgcccatca gcccaccgag cacaggtccc
      841 gcgaccatcc cgaacccgaa acaggcgctc atgaagccga agtgccgcgc gcgctcatcg
      901 ccatcagtga tatcggcaat ataagcgccg gctaccgccc cagtcgcccc ggtgatgccg
      961 gccacgatcc gcccgatata gagaacccaa aggaaaggcg ctgtcgccat gatggcgtag
     1021 tcgacagtgg cgccggccag cgagacgagc aagattggcc gccgcccgaa acgatccgac
     1081 agcgcgccca gcacaggtgc gcaggcaaat tgcaccaacg catacagcgc cagcagaatg
     1141 ccatagtggg cggtgacgtc gttcgagtga accagatcgc gcaggaggcc cggcagcacc
     1201 ggcataatca ggccgatgcc gacagcgtcg agcgcgacag tgctcagaat tacgatcagg
     1261 ggtatgttgg gtttcacgtc tggcctccgg accagcctcc gctggtccga ttgaacgcgc
     1321 ggattcttta tcactgataa gttggtggac atactatgtt tatcagtgat aaagtgtcaa
     1381 gcatgacaaa gttgcagccg aatacagtga tccgtgccgc cctggacctg ttgaacgagg
     1441 tcggcgtaga cggtctgacg acacgcaaac tggcggaacg gttgggggtt cagcagccgg
     1501 cgctttactg gcacttcagg aacaagcggg cgctgctcga cgcactggcc gaagccatgc
     1561 tggcggagaa tcatacgcat tcggtgccga gagccgacga cgactggcgc tcatttctga
     1621 tcgggaatgc ccgcagcttc aggcaggcgc tgctcgccta ccgcgatggc gcgcgcatcc
     1681 atgccggcac gcgaccgggc gcaccgcaga tggaaacggc cgacgcgcag cttcgcttcc
     1741 tctgcgaggc gggtttttcg gccggggacg ccgtcaatgc gctgatgaca atcagctact
     1801 tcactgttgg ggccgtgctt gaggagcagg ccggcgacag cgatgccggc gagcgcggcg
     1861 gcaccgttga acaggctccg ctctcgccgc tgttgcgggc cgcgatagac gccttcgacg
     1921 aagccggtcc ggacgcagcg ttcgagcagg gactcgcggt gattgtcgat ggattggcga
     1981 aaaggaggct cgttgtcagg aacgttgaag gaccgagaaa gggtgacgat tgactcagga
     2041 gaccattggg cccgtcgact gcagaggcct gcatgcaagc ttggcgtaat catggtcata
     2101 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2161 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2221 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2281 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2341 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2401 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2461 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2521 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2581 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2641 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2701 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2761 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2821 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2881 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2941 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3001 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3061 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3121 tcagtggaac gaaaactcac gttaagggat tttggtcatg aggcgcacac cgtggaaacg
     3181 gatgaaggca cgaacccagt tgacataagc ctgttcggtt cgtaaactgt aatgcaagta
     3241 gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc
     3301 agtggcggtt ttcatggctt gttatgactg tttttttgta cagtctatgc ctcgggcatc
     3361 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt
     3421 tacgcagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaggtggct
     3481 caagtatggg catcattcgc acatgtaggc tcggccctga ccaagtcaaa tccatgcggg
     3541 ctgctcttga tcttttcggt cgtgagttcg gcgacgtagc cacctactcc caacatcagc
     3601 cggactccga ttacctcggg aacttgctcc gtagtaagac attcatcgcg cttgctgcct
     3661 tcgaccaaga agcggttgtt ggcgctctcg cggcttacgt tctgcccaag tttgagcagc
     3721 cgcgtagtga gatctatatc tatgatctcg cagtctccgg agagcaccgg aggcagggca
     3781 ttgccaccgc gctcatcaat ctcctcaagc atgaggccaa cgcgcttggt gcttatgtga
     3841 tctacgtgca agcagattac ggtgacgatc ccgcagtggc tctctataca aagttgggca
     3901 tacgggaaga agtgatgcac tttgatatcg acccaagtac cgccacctaa caattcgttc
     3961 aagccgagat cggcttcccg gccgcggagt tgttcggtaa attgtcacaa cgccgctcat
     4021 gacattaacc tataaaaata ggcgtatcac gaggcccttt tgtctcgcgc gtttcggtga
     4081 tgacggtgaa aacctctgac acatgcagct cccggagaag gtcacagctt gtctgtaagc
     4141 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     4201 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     4261 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     4321 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     4381 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     4441 tt
//
