LOCUS       sequence-201158-        2617 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LI C-terminal module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2617)
  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 2617)
  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 D-E V5
FEATURES             Location/Qualifiers
     source          1..2617
                     /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             100..141
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             142..183
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             184..225
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(270..286)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(270..286)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(283..305)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    294..310
                     /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(318..324,325..342,343..348))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    363..384
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(501..518)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(672..1260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(752..771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1375..1380,1381..1397,1398..1403)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1592..2125
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     2212..2230
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2268..2290)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2390..2409
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(2603..2617,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(2617,1..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 aggatgggtc tcaaaggttg gaaagcccat tcccaaccca
      121 cttcttggtt tagatagcac cggaaagcct atccctaatc ctttacttgg tcttgatagt
      181 actggaaaac caattccgaa ccctctgcta ggattggatt ctacataaaa tcagagacca
      241 tgcctgcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
      301 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
      361 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
      421 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
      481 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
      541 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
      601 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
      661 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      721 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      781 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
      841 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
      901 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
      961 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1021 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1081 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1141 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1201 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1261 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1321 gggattttgg tcatgaggcg cacaccgtgg aaacggatga aggcacgaac ccagttgaca
     1381 taagcctgtt cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg caactggtcc
     1441 agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat
     1501 gactgttttt ttgtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
     1561 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagcaacg atgttacgca
     1621 gcagggcagt cgccctaaaa caaagttagg tggctcaagt atgggcatca ttcgcacatg
     1681 taggctcggc cctgaccaag tcaaatccat gcgggctgct cttgatcttt tcggtcgtga
     1741 gttcggcgac gtagccacct actcccaaca tcagccggac tccgattacc tcgggaactt
     1801 gctccgtagt aagacattca tcgcgcttgc tgccttcgac caagaagcgg ttgttggcgc
     1861 tctcgcggct tacgttctgc ccaagtttga gcagccgcgt agtgagatct atatctatga
     1921 tctcgcagtc tccggagagc accggaggca gggcattgcc accgcgctca tcaatctcct
     1981 caagcatgag gccaacgcgc ttggtgctta tgtgatctac gtgcaagcag attacggtga
     2041 cgatcccgca gtggctctct atacaaagtt gggcatacgg gaagaagtga tgcactttga
     2101 tatcgaccca agtaccgcca cctaacaatt cgttcaagcc gagatcggct tcccggccgc
     2161 ggagttgttc ggtaaattgt cacaacgccg ctcatgacat taacctataa aaataggcgt
     2221 atcacgaggc ccttttgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     2281 cagctcccgg agaaggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     2341 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     2401 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     2461 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     2521 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     2581 agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
