LOCUS       sequence-499188-        4028 bp    DNA     circular SYN 05-FEB-2026
DEFINITION  Encodes SYRAP (RAP2.12-GAL4STE12), Gateway compatible as Donor 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4028)
  AUTHORS   Lavilla-Puerta M, He Y, Piccinini L, Di Paco L, Papachristodoulou A,
            Licausi F, Giuntoli B
  TITLE     A synthetic ERFVII-dependent circuit in yeast sheds light on the 
            regulation of early hypoxic responses of plants
  JOURNAL   bioRxiv 2025.09.01.673502
REFERENCE   2  (bases 1 to 4028)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 5, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.09.01.673502"
COMMENT     Sequence Label: SYRAP_pDONR201
FEATURES             Location/Qualifiers
     source          1..4028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(442..1251)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1165..1184
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1243..1262
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1374..1473
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1730..2170)
                     /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"
     protein_bind    complement(3247..3346)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3399..3485
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3577..3604
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(join(3739..4028,1..299))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3819..3838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
       61 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
      121 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
      181 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
      241 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
      301 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
      361 ggattttggt catgagcttg cgccgtcccg tcaagtcagc gtaatgctct gccagtgtta
      421 caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt
      481 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
      541 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
      601 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
      661 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
      721 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
      781 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
      841 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
      901 attttcacct gaatcaggat attcttctaa tacctggaat gctgtttttc cggggatcgc
      961 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     1021 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     1081 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca agcgatagat
     1141 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     1201 catgttggaa tttaatcgcg gcctcgacgt ttcccgttga atatggctca taacacccct
     1261 tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg
     1321 tgcaatgtaa catcagagat tttgagacac gggccagagc tgcagctgga tggcaaataa
     1381 tgattttatt ttgactgata gtgacctgtt cgttgcaaca aattgataag caatgctttc
     1441 ttataatgcc aactttgtac aagaaagctg ggtgttagaa accgttgttg tttggatcgg
     1501 ttttcaagaa tggcaaacca tcagacatat catcctcttc agaccaatca aaaccatgca
     1561 atgcatctct tggcatgaag tctaatggga tagtagagtt atcatgatga gcagcagaat
     1621 caatagctgg agtaaaagct tgtggtggaa atggatcgaa ggcattttct tcagttggat
     1681 attcaacaga gactgggaag taatccaatg gaaaatcttc ttcatctcta gacacggtca
     1741 actgtctttg acctttgtta gaagattctt cagagctgga ggtagcagaa attctatgtt
     1801 gtctcaaggt caatggcata tcagtttcaa cagaagccaa tctatcagta acggcatcct
     1861 tgttaacgtt atcttgaacg aacaaaccag tcaacaaagc cttgatatct tgcaaggaat
     1921 ccatcttcaa gatcatgtcc agatcttctc ttgggaaaat caacaagaac agctgctcca
     1981 atctttccaa cctagattca acttcggtca aatgtgctct agtcaatgga gatctcttag
     2041 tctttggaga gtatctacat tcccagttgt tcttcaaaca cttagcacac tttggctttt
     2101 ctttggagca cttcaatttt ttcagcctgc agatatcgca agcttgttca atagaggaca
     2161 acagcttcat gaagactcct ccaatcatgg aatgaatttc atcaatactc cataggtcca
     2221 tagggtttgc accattgtcc tgagtcgtta cagcatcttc gttgaggaaa tccagtgaag
     2281 cgtcccattc agtgttgttg taaggtgtct cgaaatccat agacttgagc ttcttagctg
     2341 gattggcttc ctcaaagaac agagctgagt tgttgttgtt gataaccgca gaagagatgt
     2401 cgggagttat cggagcttga tcgctccaac caaactccga acaatcgaaa gaattactac
     2461 cctggtcaga gctgaaatac tgatacccat tacatccagc atcaacggag tttgtcatcc
     2521 caaactggtt gttgttgttg ttgctcactt ggtgtttctc ctccatgaaa cacatatttt
     2581 caaaggagat gttcgagttc tgaaccaaag ctggacttgg gttagggtta ggtttagcca
     2641 ctggtttctg aagattagcc ttcacagagc gtttctgaga attagccttc atgttttctt
     2701 cagggaaatt caccttagct ttagatccac ggattctccg cgctgcagca tcgtaagctc
     2761 tagcagcttc ctcagctgtc ttgaacgttc caagccagat tctagcacct tcccttggat
     2821 cacgtatctc agcagcccat tttccccaag gacgttgcct aatccctcgg tactgattct
     2881 tcctcttcct atttgcagat ttctcagcgt ccccatccaa gccagtaact ttcttaccaa
     2941 aaactgaacc ttccgcagcg gcggagacgg cgggttttgg agtcgaagtg aaaacgaatg
     3001 gtttcacatc ggcgaaaaca tcaccgacgt cgaaatcatc atcgcaatcg atagacgaat
     3061 catctttgaa accttggaaa tcagcttcga actcagcgtc aaaatcgaag aaattcgaac
     3121 gattcttcga gcttttcttc gatcctttca aattcttctt cagatccggc caaataaact
     3181 cgctagtaac acggcgagac ctcggcggtg gaatgaaatc ggatattata gctcctccac
     3241 acatggagcc tgcttttttg tacaaagttg gcattataaa aaagcattgc tcatcaattt
     3301 gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttggggc ccgagatcca
     3361 tgctagcgtt aacgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     3421 cggaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     3481 caaatccgcc gggagcggat ttgaacgttg tgaagcaacg gcccggaggg tggcgggcag
     3541 gacgcccgcc ataaactgcc aggcatcaaa ctaagcagaa ggccatcctg acggatggcc
     3601 tttttgcgtt tctacaaact cttcctggct agcggtaata cggttatcca cagaatcagg
     3661 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3721 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3781 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3841 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3901 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3961 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4021 ctgcgcct
//
