LOCUS       sequence-371931-        5825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enables expression of OsTIR1(F74G) from the TEF2 promoter, used to 
            implement the AID2 system in Komagataella phaffii (original name 
            pLB219).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5825)
  AUTHORS   Lehmayer L, Bernauer L, Emmerstorfer-Augustin A
  TITLE     Applying the auxin-based degron system for the inducible, reversible
            and complete protein degradation in Komagataella phaffii.
  JOURNAL   iScience. 2022 Aug 6;25(9):104888. doi: 10.1016/j.isci.2022.104888. 
            eCollection 2022 Sep 16.
  PUBMED    36043049
REFERENCE   2  (bases 1 to 5825)
  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: "iScience.";
            date: "2022-08-6"; volume: "25(9):104888. doi"; pages: " 
            10.1016/j.isci.2022.104888. eCollection 2022 Sep 16"
COMMENT     Sequence Label: pPpT4- TEF2prom-TIR1F74G-FLAG
FEATURES             Location/Qualifiers
     source          1..5825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1653..3377
                     /codon_start=1
                     /product="rice transport inhibitor response 1 protein, 
                     mutated for use in the AID2 system of inducible degradation
                     (Yesbolatova et al., 2020)"
                     /label=OsTIR1(F74G)
                     /note="F-box protein that mediates auxin-dependent 
                     interaction of a ubiquitin ligase with an auxin-inducible 
                     degron (Tan et al., 2007)"
                     /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV
                     GNCYAVRAGRVAARFPNVRALTVKGKPHGADFNLVPPDWGGYAGPWIEAAARGCHGLEE
                     LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV
                     EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL
                     AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP
                     LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR
                     VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL
                     CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI
                     AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG
                     CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL"
     CDS             3378..3401
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      3413..3659
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     rep_origin      5195..5783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5684..5703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgta tttaaattgc cagtgtatgt gcacttatag aggatgtaca
       61 aatggtcaga gtttccccaa agagcagatg gaaaagcaat tagattttcg ttactgaatc
      121 aggcattcaa aaagaaacaa taagtcgcaa caatgatgtt aatctcgaat attgaagtaa
      181 atgcttccag caataatgtg acagttgctc gaatctagaa cggtattaac agaatagacc
      241 gagactctcc tatttcagcg tcctaaactc tatcttggga ccaaggaaga aagctacgca
      301 cttaagaata ggaccatgca taccaaaaca atagacctga cctgatacgc tatgcactgc
      361 aaatcgaaga aaaaagcaat tcattcggaa caaaacgtca gagcgggttt tctaatagaa
      421 tttgcgtaat atataaacct aaataccata attgaggtgt ttcttttctc tgataaaagt
      481 acaacgaagt gctcttgtca taccgcaaca aagatgttga acacttcgta tcagtttccc
      541 aaagatgttg caagatacga cgtggcagat gctgtcaatg agcaacttca agctaccttt
      601 ttgaattcaa aggttcaaca tgagcccggt tcaagatatc aacaccaaga gggaatcaac
      661 tcatcaccag agagaaaaaa ttcagtcttt tcatctgctg atatatctac cacatcgtca
      721 cctacttctc cgaagtctaa agccaattct caatttgcta gagactctgt cgacagttct
      781 atctacccga ggaaaccgat acataggaga tctaatacag aggtatcatc caatgattcc
      841 gaaaaagatt tcagtttgga ctcccttttt aaagcacagt atcgggatac aaagctgaaa
      901 ccctccccat ttcaagcttc tagtggtagg aattctgtac cggtttacag aaggacgact
      961 cttgatgcgc caaccacagt gacaatagac atagaggaaa tgacaaaagg attatgcgag
     1021 gatgctgctg gagacgattc aaagtttagt ttagaaaggt cctccattta tgctgataga
     1081 atactagata cccgtgaact ttgtctcagg agatccgcat cagacgaagg atgttccgac
     1141 ctgcaaataa tcgaagaaga gacccctagg ccgtacgaca gtcagttagt agatatttat
     1201 accattctgc gagaaggtcc cctgcaggat aactgtcgcc tcttttatct gccgcactgc
     1261 atgaggtgtc cccttagtgg gaaagagtac tgagccaacc ctggaggaca gcaagggaaa
     1321 aatacctaca acttgcttca taatggtcgt aaaaacaatc cttgtcggat ataagtgttg
     1381 tagactgtcc cttatcctct gcgatgttct tcctctcaaa gtttgcgatt tctctctatc
     1441 agaattgcca tcaagagact caggactaat ttcgcagtcc cacacgcact cgtacatgat
     1501 tggctgaaat ttccctaaag aatttctttt tcacgaaaat ttttttttta cacaagattt
     1561 tcagcagata taaaatggag agcaggacct ccgctgtgac tcttcttttt tttcttttat
     1621 tctcactaca tacattttag ttattcgcca acatgactta ttttcctgaa gaagttgtag
     1681 aacacatatt ttccttttta cctgctcaaa gagatagaaa caccgtctcc ttggtctgta
     1741 aggtttggta cgaaatcgaa agattgtcta gaagaggtgt ttttgttggt aattgttacg
     1801 cagttagagc tggtagagtt gctgcaagat tcccaaacgt tagagcattg actgttaaag
     1861 gtaaacctca tggtgctgat ttcaatttgg ttccacctga ttggggtggt tatgctggtc
     1921 catggattga agctgcagct agaggttgtc acggtttgga agaattgaga atgaagagaa
     1981 tggttgtttc tgatgaatca ttggaattgt tagcaagatc ttttcctaga ttcagagctt
     2041 tggttttgat ctcatgtgaa ggtttttcca ccgatggttt agcagctgtt gcttcacatt
     2101 gtaagttgtt gagagaattg gatttgcaag aaaacgaagt tgaagataga ggtccaagat
     2161 ggttatcttg ttttcctgat tcctgtacaa gtttggtttc tttgaacttc gcatgtatca
     2221 agggtgaagt taacgctggt agtttggaaa gattagtttc tagatcacca aatttgagat
     2281 ctttgagatt gaacagatcc gttagtgttg ataccttggc aaagatcttg ttgagaactc
     2341 ctaatttgga agatttgggt actggtaact tgacagatga tttccaaaca gaatcatact
     2401 tcaagttgac ctccgctttg gaaaagtgta agatgttgag atctttgtca ggtttttggg
     2461 atgcaagtcc agtttgtttg tcttttatat atcctttgtg tgctcaattg actggtttga
     2521 atttgtctta cgcaccaact ttggatgcta gtgatttgac aaagatgatc tctagatgtg
     2581 ttaagttgca aagattgtgg gttttggatt gtatctcaga taaaggtttg caagttgttg
     2641 cttcttcttg taaggatttg caagaattga gagtttttcc aagtgatttc tatgttgcag
     2701 gttactctgc tgttacagaa gaaggtttag ttgcagtttc attgggttgt cctaagttga
     2761 actccttgtt gtacttctgt catcaaatga ccaacgcagc tttggttact gttgctaaaa
     2821 attgtccaaa cttcactaga ttcagattgt gtatcttgga accaggtaaa cctgatgttg
     2881 ttacatcaca acctttagat gaaggttttg gtgcaatcgt tagagagtgt aagggtttgc
     2941 aaagattgtc catcagtggt ttgttgacag ataaggtttt tatgtacatc ggtaaatacg
     3001 ctaagcaatt ggaaatgttg tcaatcgcat tcgctggtga ctccgataag ggtatgatgc
     3061 atgttatgaa cggttgtaag aacttaagaa agttggaaat cagagattct ccatttggtg
     3121 acgcagcttt gttgggtaat ttcgcaagat atgaaacaat gagatctttg tggatgtcca
     3181 gttgtaacgt taccttgaaa ggttgtcaag ttttagcttc aaagatgcca atgttgaacg
     3241 ttgaagttat taacgaaaga gatggttcca atgaaatgga agaaaaccat ggtgacttgc
     3301 ctaaagttga aaagttatac gtctacagaa caaccgcagg tgccagagat gatgctccta
     3361 acttcgtcaa gattttagat tataaagatg acgatgacaa gtgagcggcc gctcaagagg
     3421 atgtcagaat gccatttgcc tgagagatgc aggcttcatt tttgatactt ttttatttgt
     3481 aacctatata gtataggatt ttttttgtca ttttgtttct tctcgtacga gcttgctcct
     3541 gatcagccta tctcgcagca gatgaatatc ttgtggtagg ggtttgggaa aatcattcga
     3601 gtttgatgtt tttcttggta tttcccactc ctcttcagag tacagaagat taagtgagac
     3661 cttcgtttgt gcggatcctt cagtaatgtc ttgtttcttt tgttgcagtg gtgagccatt
     3721 ttgacttcgt gaaagtttct ttagaatagt tgtttccaga ggccaaacat tccacccgta
     3781 gtaaagtgca agcgtaggaa gaccaagact ggcataaatc aggtataagt gtcgagcact
     3841 ggcaggtgat cttctgaaag tttctactag cagataagat ccagtagtca tgcatatggc
     3901 aacaatgtac cgtgtggatc taagaacgcg tcctactaac cttcgcattc gttggtccag
     3961 tttgttgtta tcgatcaacg tgacaaggtt gtcgattccg cgtaagcatg catacccaag
     4021 gacgcctgtt gcaattccaa gtgagccagt tccaacaatc tttgtaatat tagagcactt
     4081 cattgtgttg cgcttgaaag taaaatgcga acaaattaag agataatctc gaaaccgcga
     4141 cttcaaacgc caatatgatg tgcggcacac aataagcgtt catatccgct gggtgacttt
     4201 ctcgctttaa aaaattatcc gaaaaaattt tctagagtgt tgttacttta tacttccggc
     4261 tcgtataata cgacaaggtg taaggaggac taaaccatgg ctaaactcac ctctgctgtt
     4321 ccagtcctga ctgctcgtga tgttgctggt gctgttgagt tctggactga tagactcggt
     4381 ttctcccgtg acttcgtaga ggacgacttt gccggtgttg tacgtgacga cgttaccctg
     4441 ttcatctccg cagttcagga ccaggttgtg ccagacaaca ctctggcatg ggtatgggtt
     4501 cgtggtctgg acgaactgta cgctgagtgg tctgaggtcg tgtctaccaa cttccgtgat
     4561 gcatctggtc cagctatgac cgagatcggt gaacagccct ggggtcgtga gtttgcactg
     4621 cgtgatccag ctggtaactg cgtgcatttc gtcgcagaag agcaggacta acaattgaca
     4681 ccttacgatt atttagagag tatttattag ttttattgta tgtatacgga tgttttatta
     4741 tctatttatg cccttatatt ctgtaactat ccaaaagtcc tatcttatca agccagcaat
     4801 ctatgtccgc gaacgtcaac taaaaataag ctttttatgc tcttctctct ttttttccct
     4861 tcggtataat tataccttgc atccacagat tctcctgcca aattttgcat aatcctttac
     4921 aacatggcta tatgggagca cttagcgccc tccaaaaccc atattgccta cgcatgtata
     4981 ggtgtttttt ccacaatatt ttctctgtgc tctcttttta ttaaagagaa gctctatatc
     5041 ggagaagctt ctgtggccgt tatattcggc cttatcgtgg gaccacattg cctgaattgg
     5101 tttgccccgg aagattgggg aaacttggat ctgattacct tagctgcagg taccactgag
     5161 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5221 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5281 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5341 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5401 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5461 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5521 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5581 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5641 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5701 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5761 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5821 tttgc
//
