LOCUS       sequence-418451-        3538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate LI D-E module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3538)
  AUTHORS   Tan H, Zhou Y, Dinius E, Lozano-Duran R
  TITLE     The Ti-TAN plasmid toolbox for TurboID-based proximity labeling 
            assays in Nicotiana benthamiana.
  JOURNAL   J Integr Plant Biol. 2024 Jan 31. doi: 10.1111/jipb.13610.
  PUBMED    38294100
REFERENCE   2  (bases 1 to 3538)
  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: "J Integr 
            Plant Biol. 2024 Jan 31. doi: 10.1111/jipb.13610."
COMMENT     Sequence Label: LI D-E TurboID
FEATURES             Location/Qualifiers
     source          1..3538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     466..484
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(522..544)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     644..663
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     857..879
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     871..888
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     872..888
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(950..1906)
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             complement(1913..1939)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1943..1969)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1973..1999)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2062..2078)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2062..2078)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2075..2097)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2086..2102
                     /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(2110..2116,2117..2134,2135..2140))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2155..2176
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2293..2310)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2464..3052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2544..2563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(3167..3172,3173..3189,3190..3195)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(3384..3538,1..379)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 cgacgtagcc acctactccc aacatcagcc ggactccgat tacctcggga acttgctccg
       61 tagtaagaca ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc
      121 ggcttacgtt ctgcccaagt ttgagcagcc gcgtagtgag atctatatct atgatctcgc
      181 agtctccgga gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca
      241 tgaggccaac gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc
      301 cgcagtggct ctctatacaa agttgggcat acgggaagaa gtgatgcact ttgatatcga
      361 cccaagtacc gccacctaac aattcgttca agccgagatc ggcttcccgg ccgcggagtt
      421 gttcggtaaa ttgtcacaac gccgctcatg acattaacct ataaaaatag gcgtatcacg
      481 aggccctttt gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
      541 ccggagaagg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
      601 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
      661 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
      721 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
      781 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
      841 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcgagctc
      901 ggtacctcgc gaatgcatct agatatcgga tcccaggtct cagattctac ttttcggcag
      961 accgcagact gatttctccg cccatccagg gtttgataac tccgtcctgt tccagcagca
     1021 gggctccctg tttatcaatc ccccggctaa tcccgaaaat ctctttgtcc ccaatgatca
     1081 gcttcacggg tctattgatg aagttatcca gcttctccca ccgtggcaga tatggagcca
     1141 ggccttcctg ctcgaacagt tccagggctg cccgcagctc tcggatcagc gtagcggcca
     1201 gagtattcct gtccaggtta atccctgctt cctgcagtgt gatccagccc tgattgacca
     1261 cgctttcctc cacgcgcctc atagccacgt taatcccggc gccaatgacg atctgtgcag
     1321 catcgcctgt tattccggcc agctccacca ggatgcctgc cagctttcta tcctgcagat
     1381 acaggtcatt gggccatttg actcgcacct tgtctgctcc cagctttcgc agcgcttctg
     1441 ccatgacaat tccgatgacc gggcccaggc cgattgctgc tggtccccgc ttcaggcgcc
     1501 aaaacatact caggtacagg ttagctccaa aaggagagaa ccatttcctc cctctgcttc
     1561 ctctccctgc ctgctgatat tctgcaatgc aagcatcccc actcttcagc tcgccgattc
     1621 gatccagcag gtactgattg gtggagtcta ccacaggcag gactgccacg ctcccgccgt
     1681 ccagctgtcc cagaatctgt ttagcgttca gcagcgggat aggctcgggc agagagtagc
     1741 cctttccggg cacggtgaac acgtccacgc cccagtcgcg cagagtctga atgtgcttgt
     1801 tgatagcggc cctggacatg cccagggttt ctcccagctg ttcgccacta tggaactcgc
     1861 cattagccag gagagcgatc agcttcagag gcacagtatt gtctttgcta gctgcgtagt
     1921 ctgggacgtc ataggggtac ccggcatagt cgggcacgtc gtagggatag ccagcgtaat
     1981 ccggaacatc atacgggtat gcctttgaga cccggggccc gtcgactgca gaggcctgca
     2041 tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2101 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     2161 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2221 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2281 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2341 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2401 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2461 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2521 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2581 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2641 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2701 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2761 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2821 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2881 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2941 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3001 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3061 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3121 ggtcatgagg cgcacaccgt ggaaacggat gaaggcacga acccagttga cataagcctg
     3181 ttcggttcgt aaactgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt
     3241 gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt
     3301 ttttgtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg
     3361 tttgatgtta tggagcagca acgatgttac gcagcagcaa cgatgttacg cagcagggca
     3421 gtcgccctaa aacaaagtta ggtggctcaa gtatgggcat cattcgcaca tgtaggctcg
     3481 gccctgacca agtcaaatcc atgcgggctg ctcttgatct tttcggtcgt gagttcgg
//
