LOCUS       sequence-401503-        4120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GAL4-VP16 transcription factor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4120)
  AUTHORS   Chen J, Luo M, Hands P, Rolland V, Zhang J, Li Z, Outram M, Dodds P,
            Ayliffe M
  TITLE     A split GAL4 RUBY assay for visual in planta detection of 
            protein-protein interactions.
  JOURNAL   Plant J. 2023 Jun;114(6):1209-1226. doi: 10.1111/tpj.16234. Epub 
            2023 Apr 26.
  PUBMED    37323061
REFERENCE   2  (bases 1 to 4120)
  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"; doi: "10.1111/tpj.16234";
            journalName: "Plant J"; date: "2023-06"; volume: "114"; issue: "6"; 
            pages: "1209-1226"
COMMENT     Sequence Label: GAL4BD-VP16/pDON207
FEATURES             Location/Qualifiers
     source          1..4120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(89..116)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(208..294)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    347..446
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             449..889
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             911..1144
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     CDS             1145..1165
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1172..1192
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(1197..1296)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1408..1427)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1486..1505)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(2079..2612)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2801..2806,2807..2823,2824..2829))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3486..4074
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3975..3994
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
       61 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
      121 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
      181 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
      241 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
      301 ggcagttccc tactctcgcg ttaacgctag catggatctc gggccccaaa taatgatttt
      361 attttgactg atagtgacct gttcgttgca acaaattgat gagcaatgct tttttataat
      421 gccaagtttg tacaaaaaag caggcttaat gaagctactg tcttctatcg aacaagcatg
      481 cgatatttgc cgacttaaaa agctcaagtg ctccaaagaa aaaccgaagt gcgccaagtg
      541 tctgaagaac aactgggagt gtcgctactc tcccaaaacc aaaaggtctc cgctgactag
      601 ggcacatctg acagaagtgg aatcaaggct agaaagactg gaacagctat ttctactgat
      661 ttttcctcga gaagaccttg acatgatttt gaaaatggat tctttacagg atataaaagc
      721 attgttaaca ggattatttg tacaagataa tgtgaataaa gatgccgtca cagatagatt
      781 ggcttcagtg gagactgata tgcctctaac attgagacag catagaataa gtgcgacatc
      841 atcatcggaa gagagtagta acaaaggtca aagacagttg actgtatcgc cggaattccc
      901 ggggatctgg gcccccccga ccgatgtcag cctgggggac gagctccact tagacggcga
      961 ggacgtggcg atggcgcatg ccgacgcgct agacgatttc gatctggaca tgttggggga
     1021 cggggattcc ccgggtccgg gatttacccc ccacgactcc gccccctacg gcgctctgga
     1081 tatggccgac ttcgagtttg agcagatgtt taccgatgcc cttggaattg acgagtacgg
     1141 tgggcctaag aagaagcgga aagtggagga tcccaagaag aagcgcaaag tctagaaccc
     1201 agctttcttg tacaaagtgg gcattataag aaagcattgc ttatcaattt gttgcaacga
     1261 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgcag ctctggcccg
     1321 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     1381 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     1441 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     1501 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
     1561 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     1621 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     1681 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
     1741 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     1801 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     1861 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     1921 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     1981 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     2041 aattaggccg ggaagccgat ctcggcttga acgaattgtt aggtggcggt acttgggtcg
     2101 atatcaaagt gcatcacttc ttcccgtatg cccaactttg tatagagagc cactgcggga
     2161 tcgtcaccgt aatctgcttg cacgtagatc acataagcac caagcgcgtt ggcctcatgc
     2221 ttgaggagat tgatgagcgc ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg
     2281 agatcataga tatagatctc actacgcggc tgctcaaact tgggcagaac gtaagccgcg
     2341 agagcgccaa caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg
     2401 agcaagttcc cgaggtaatc ggagtccggc tgatgttggg agtaggtggc tacgtctccg
     2461 aactcacgac cgaaaagatc aagagcagcc cgcatggatt tgacttggtc agggccgagc
     2521 ctacatgtgc gaatgatgcc catacttgag ccacctaact ttgttttagg gcgactgccc
     2581 tgctgcgtaa catcgttgct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg
     2641 acccacggcg taacgcgctt gctgcttgga tgcccgaggc atagactgta caaaaaaaca
     2701 gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt
     2761 tctggaccag ttgcgtgagc gcatacgcta cttgcattac agtttacgaa ccgaacaggc
     2821 ttatgtcaac tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc cggcaacctt
     2881 gggcagcagc gaagtcgagg catttctgtc ctggctggcg aacgagcgca aggtttcggt
     2941 ctccacgcat cgtcaggcat tggcggcctt gctgttcttc tacggcaagg tgctgtgcac
     3001 ggatctgccc tggcttcagg agatcggaag acctcggccg tcgcggcgct tgccggtggt
     3061 gctgaccccg gatgaagtgg ttcgcatcct cggttttctg gaaggcgagc atcgtttgtt
     3121 cgcccaggac tctagctata gttctagtgg ttggctacta ataggttgta ttgatgttgg
     3181 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3241 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3301 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3361 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     3421 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3481 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3541 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3601 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3661 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3721 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3781 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3841 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3901 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3961 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4021 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4081 agcaacgcgg cctttttacg gttcctggcc ttttgctggc
//
