LOCUS       sequence-371930-        5860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enables expression of OsTIR1(F74G) from the HTA1 promoter, used to 
            implement the AID2 system in Komagataella phaffii (original name 
            pLB220).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5860)
  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 5860)
  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- HTA1prom-TIR1 F74G -FLAG
FEATURES             Location/Qualifiers
     source          1..5860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1688..3412
                     /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             3413..3436
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      3448..3694
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     rep_origin      5230..5818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5719..5738
                     /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 cctgcaggga ggactctcgt ttcctatgat accttatgga
     1261 atgggacagt acgcgctaat ttgtctcgtt ctggatagac tgctctgttt ttcgttacat
     1321 acgagtggga agtgcgaact tttgtaccag agtgcgaaaa aagtggtggc aatcctgacg
     1381 tttgtaccaa gcgcgacggc acattattgg tcagtttgtg cattttagtt tttcttgaaa
     1441 acatctcgaa acataacaac taggacacca attctacagt gaaaaccatt cagtggaatg
     1501 tgaacccaca taaatcttgc agcaataata caacaattta tggctcctgc attctgattg
     1561 gctgagcgtt tattgccatc gtgactgtcc aatttaggac acaccctttc ctcagaatat
     1621 ataaactggt gataattcct tcgttctgag ttccatctca tactcaaact atattaaaac
     1681 tacaacaatg acttattttc ctgaagaagt tgtagaacac atattttcct ttttacctgc
     1741 tcaaagagat agaaacaccg tctccttggt ctgtaaggtt tggtacgaaa tcgaaagatt
     1801 gtctagaaga ggtgtttttg ttggtaattg ttacgcagtt agagctggta gagttgctgc
     1861 aagattccca aacgttagag cattgactgt taaaggtaaa cctcatggtg ctgatttcaa
     1921 tttggttcca cctgattggg gtggttatgc tggtccatgg attgaagctg cagctagagg
     1981 ttgtcacggt ttggaagaat tgagaatgaa gagaatggtt gtttctgatg aatcattgga
     2041 attgttagca agatcttttc ctagattcag agctttggtt ttgatctcat gtgaaggttt
     2101 ttccaccgat ggtttagcag ctgttgcttc acattgtaag ttgttgagag aattggattt
     2161 gcaagaaaac gaagttgaag atagaggtcc aagatggtta tcttgttttc ctgattcctg
     2221 tacaagtttg gtttctttga acttcgcatg tatcaagggt gaagttaacg ctggtagttt
     2281 ggaaagatta gtttctagat caccaaattt gagatctttg agattgaaca gatccgttag
     2341 tgttgatacc ttggcaaaga tcttgttgag aactcctaat ttggaagatt tgggtactgg
     2401 taacttgaca gatgatttcc aaacagaatc atacttcaag ttgacctccg ctttggaaaa
     2461 gtgtaagatg ttgagatctt tgtcaggttt ttgggatgca agtccagttt gtttgtcttt
     2521 tatatatcct ttgtgtgctc aattgactgg tttgaatttg tcttacgcac caactttgga
     2581 tgctagtgat ttgacaaaga tgatctctag atgtgttaag ttgcaaagat tgtgggtttt
     2641 ggattgtatc tcagataaag gtttgcaagt tgttgcttct tcttgtaagg atttgcaaga
     2701 attgagagtt tttccaagtg atttctatgt tgcaggttac tctgctgtta cagaagaagg
     2761 tttagttgca gtttcattgg gttgtcctaa gttgaactcc ttgttgtact tctgtcatca
     2821 aatgaccaac gcagctttgg ttactgttgc taaaaattgt ccaaacttca ctagattcag
     2881 attgtgtatc ttggaaccag gtaaacctga tgttgttaca tcacaacctt tagatgaagg
     2941 ttttggtgca atcgttagag agtgtaaggg tttgcaaaga ttgtccatca gtggtttgtt
     3001 gacagataag gtttttatgt acatcggtaa atacgctaag caattggaaa tgttgtcaat
     3061 cgcattcgct ggtgactccg ataagggtat gatgcatgtt atgaacggtt gtaagaactt
     3121 aagaaagttg gaaatcagag attctccatt tggtgacgca gctttgttgg gtaatttcgc
     3181 aagatatgaa acaatgagat ctttgtggat gtccagttgt aacgttacct tgaaaggttg
     3241 tcaagtttta gcttcaaaga tgccaatgtt gaacgttgaa gttattaacg aaagagatgg
     3301 ttccaatgaa atggaagaaa accatggtga cttgcctaaa gttgaaaagt tatacgtcta
     3361 cagaacaacc gcaggtgcca gagatgatgc tcctaacttc gtcaagattt tagattataa
     3421 agatgacgat gacaagtgag cggccgctca agaggatgtc agaatgccat ttgcctgaga
     3481 gatgcaggct tcatttttga tactttttta tttgtaacct atatagtata ggattttttt
     3541 tgtcattttg tttcttctcg tacgagcttg ctcctgatca gcctatctcg cagcagatga
     3601 atatcttgtg gtaggggttt gggaaaatca ttcgagtttg atgtttttct tggtatttcc
     3661 cactcctctt cagagtacag aagattaagt gagaccttcg tttgtgcgga tccttcagta
     3721 atgtcttgtt tcttttgttg cagtggtgag ccattttgac ttcgtgaaag tttctttaga
     3781 atagttgttt ccagaggcca aacattccac ccgtagtaaa gtgcaagcgt aggaagacca
     3841 agactggcat aaatcaggta taagtgtcga gcactggcag gtgatcttct gaaagtttct
     3901 actagcagat aagatccagt agtcatgcat atggcaacaa tgtaccgtgt ggatctaaga
     3961 acgcgtccta ctaaccttcg cattcgttgg tccagtttgt tgttatcgat caacgtgaca
     4021 aggttgtcga ttccgcgtaa gcatgcatac ccaaggacgc ctgttgcaat tccaagtgag
     4081 ccagttccaa caatctttgt aatattagag cacttcattg tgttgcgctt gaaagtaaaa
     4141 tgcgaacaaa ttaagagata atctcgaaac cgcgacttca aacgccaata tgatgtgcgg
     4201 cacacaataa gcgttcatat ccgctgggtg actttctcgc tttaaaaaat tatccgaaaa
     4261 aattttctag agtgttgtta ctttatactt ccggctcgta taatacgaca aggtgtaagg
     4321 aggactaaac catggctaaa ctcacctctg ctgttccagt cctgactgct cgtgatgttg
     4381 ctggtgctgt tgagttctgg actgatagac tcggtttctc ccgtgacttc gtagaggacg
     4441 actttgccgg tgttgtacgt gacgacgtta ccctgttcat ctccgcagtt caggaccagg
     4501 ttgtgccaga caacactctg gcatgggtat gggttcgtgg tctggacgaa ctgtacgctg
     4561 agtggtctga ggtcgtgtct accaacttcc gtgatgcatc tggtccagct atgaccgaga
     4621 tcggtgaaca gccctggggt cgtgagtttg cactgcgtga tccagctggt aactgcgtgc
     4681 atttcgtcgc agaagagcag gactaacaat tgacacctta cgattattta gagagtattt
     4741 attagtttta ttgtatgtat acggatgttt tattatctat ttatgccctt atattctgta
     4801 actatccaaa agtcctatct tatcaagcca gcaatctatg tccgcgaacg tcaactaaaa
     4861 ataagctttt tatgctcttc tctctttttt tcccttcggt ataattatac cttgcatcca
     4921 cagattctcc tgccaaattt tgcataatcc tttacaacat ggctatatgg gagcacttag
     4981 cgccctccaa aacccatatt gcctacgcat gtataggtgt tttttccaca atattttctc
     5041 tgtgctctct ttttattaaa gagaagctct atatcggaga agcttctgtg gccgttatat
     5101 tcggccttat cgtgggacca cattgcctga attggtttgc cccggaagat tggggaaact
     5161 tggatctgat taccttagct gcaggtacca ctgagcgtca gaccccgtag aaaagatcaa
     5221 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5281 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5341 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5401 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5461 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5521 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5581 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5641 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5701 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5761 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5821 cgccagcaac gcggcctttt tacggttcct ggccttttgc
//
