LOCUS       sequence-373205-        6401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enables expression of OsTIR1 from the PGK1 promoter, used to 
            implement the AID system in Komagataella phaffii (original name 
            pAEA454).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6401)
  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 6401)
  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- PGK1prom-TIR1-FLAG
FEATURES             Location/Qualifiers
     source          1..6401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2229..3953
                     /codon_start=1
                     /product="rice transport inhibitor response 1 protein"
                     /label=OsTIR1
                     /note="F-box protein that mediates auxin-dependent 
                     interaction of a ubiquitin ligase with an auxin-inducible 
                     degron (Tan et al., 2007)"
                     /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV
                     GNCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEE
                     LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV
                     EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL
                     AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP
                     LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR
                     VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL
                     CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI
                     AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG
                     CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL"
     CDS             3954..3977
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      3989..4235
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     rep_origin      5771..6359
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6260..6279
                     /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 cctgcaggaa gttggtaccc agccgatcac gcctgctctg
     1261 agtttggctg gagcagcaaa tctcatgata accgaggttt aaattaaggt acataacaaa
     1321 aattcaatgt tcaaagacgc acataccaag acttactaat cgcagaatgt tggtgcagta
     1381 tttgtcgtaa gccaaaacca tcgatgttga cttcctaatt cagtctttaa accgcaaaag
     1441 gattctgatt cgcagatggc ctgatctcca aactcaggct ggggctctaa ctcgagcaag
     1501 tgtcctatgc tgtaggccgc agcccttttg gttcgacgac gtgcgtggtt atgagacgct
     1561 cggctgtttt gcgctaagct ggccgtatcg agtaaattct acaggcacct gcgaggcaag
     1621 caatctacta atgtttattt ttcgtccaac ctaattgtgg tttcaaagcg ctatcaggtg
     1681 gggggtaaga ggaatgtgag tggaaagcga aaataactgg cagctggggt cagatcccgt
     1741 gatgccacct cttgtggtat tttgaaacgc gtgttgcgat tggccgcgag aacggaaagg
     1801 aatatattta ctgccgatcg cattttggcc tcaaataaat cttgagcttt tggacataga
     1861 ttatatgttc tttcttggaa gctctttcag ctaatagtga agtgtttcct actaaggatc
     1921 gcctccaaac gttccaacta cgggcggagg ttgcaaagaa aacgggtctc tcagcgaatt
     1981 gttctcatcc atgagtgagt cctctccgtc ctttcctcgc gcctggcaat aaagcctcct
     2041 tcggaggagc tccgtctaga gaataattgc tgcctttatg actttcggac tagcgccaac
     2101 cacgaaccac accaccacac catcactgtc acccgtcata gttcatccct ctctccttat
     2161 aaagcatcta ataggttcca caattgtttg ccaaaaaatc tcttagcata gcccaattga
     2221 ttacgaaaat gacttatttt cctgaagaag ttgtagaaca catattttcc tttttacctg
     2281 ctcaaagaga tagaaacacc gtctccttgg tctgtaaggt ttggtacgaa atcgaaagat
     2341 tgtctagaag aggtgttttt gttggtaatt gttacgcagt tagagctggt agagttgctg
     2401 caagattccc aaacgttaga gcattgactg ttaaaggtaa acctcatttt gctgatttca
     2461 atttggttcc acctgattgg ggtggttatg ctggtccatg gattgaagct gcagctagag
     2521 gttgtcacgg tttggaagaa ttgagaatga agagaatggt tgtttctgat gaatcattgg
     2581 aattgttagc aagatctttt cctagattca gagctttggt tttgatctca tgtgaaggtt
     2641 tttccaccga tggtttagca gctgttgctt cacattgtaa gttgttgaga gaattggatt
     2701 tgcaagaaaa cgaagttgaa gatagaggtc caagatggtt atcttgtttt cctgattcct
     2761 gtacaagttt ggtttctttg aacttcgcat gtatcaaggg tgaagttaac gctggtagtt
     2821 tggaaagatt agtttctaga tcaccaaatt tgagatcttt gagattgaac agatccgtta
     2881 gtgttgatac cttggcaaag atcttgttga gaactcctaa tttggaagat ttgggtactg
     2941 gtaacttgac agatgatttc caaacagaat catacttcaa gttgacctcc gctttggaaa
     3001 agtgtaagat gttgagatct ttgtcaggtt tttgggatgc aagtccagtt tgtttgtctt
     3061 ttatatatcc tttgtgtgct caattgactg gtttgaattt gtcttacgca ccaactttgg
     3121 atgctagtga tttgacaaag atgatctcta gatgtgttaa gttgcaaaga ttgtgggttt
     3181 tggattgtat ctcagataaa ggtttgcaag ttgttgcttc ttcttgtaag gatttgcaag
     3241 aattgagagt ttttccaagt gatttctatg ttgcaggtta ctctgctgtt acagaagaag
     3301 gtttagttgc agtttcattg ggttgtccta agttgaactc cttgttgtac ttctgtcatc
     3361 aaatgaccaa cgcagctttg gttactgttg ctaaaaattg tccaaacttc actagattca
     3421 gattgtgtat cttggaacca ggtaaacctg atgttgttac atcacaacct ttagatgaag
     3481 gttttggtgc aatcgttaga gagtgtaagg gtttgcaaag attgtccatc agtggtttgt
     3541 tgacagataa ggtttttatg tacatcggta aatacgctaa gcaattggaa atgttgtcaa
     3601 tcgcattcgc tggtgactcc gataagggta tgatgcatgt tatgaacggt tgtaagaact
     3661 taagaaagtt ggaaatcaga gattctccat ttggtgacgc agctttgttg ggtaatttcg
     3721 caagatatga aacaatgaga tctttgtgga tgtccagttg taacgttacc ttgaaaggtt
     3781 gtcaagtttt agcttcaaag atgccaatgt tgaacgttga agttattaac gaaagagatg
     3841 gttccaatga aatggaagaa aaccatggtg acttgcctaa agttgaaaag ttatacgtct
     3901 acagaacaac cgcaggtgcc agagatgatg ctcctaactt cgtcaagatt ttagattata
     3961 aagatgacga tgacaagtga gcggccgctc aagaggatgt cagaatgcca tttgcctgag
     4021 agatgcaggc ttcatttttg atactttttt atttgtaacc tatatagtat aggatttttt
     4081 ttgtcatttt gtttcttctc gtacgagctt gctcctgatc agcctatctc gcagcagatg
     4141 aatatcttgt ggtaggggtt tgggaaaatc attcgagttt gatgtttttc ttggtatttc
     4201 ccactcctct tcagagtaca gaagattaag tgagaccttc gtttgtgcgg atccttcagt
     4261 aatgtcttgt ttcttttgtt gcagtggtga gccattttga cttcgtgaaa gtttctttag
     4321 aatagttgtt tccagaggcc aaacattcca cccgtagtaa agtgcaagcg taggaagacc
     4381 aagactggca taaatcaggt ataagtgtcg agcactggca ggtgatcttc tgaaagtttc
     4441 tactagcaga taagatccag tagtcatgca tatggcaaca atgtaccgtg tggatctaag
     4501 aacgcgtcct actaaccttc gcattcgttg gtccagtttg ttgttatcga tcaacgtgac
     4561 aaggttgtcg attccgcgta agcatgcata cccaaggacg cctgttgcaa ttccaagtga
     4621 gccagttcca acaatctttg taatattaga gcacttcatt gtgttgcgct tgaaagtaaa
     4681 atgcgaacaa attaagagat aatctcgaaa ccgcgacttc aaacgccaat atgatgtgcg
     4741 gcacacaata agcgttcata tccgctgggt gactttctcg ctttaaaaaa ttatccgaaa
     4801 aaattttcta gagtgttgtt actttatact tccggctcgt ataatacgac aaggtgtaag
     4861 gaggactaaa ccatggctaa actcacctct gctgttccag tcctgactgc tcgtgatgtt
     4921 gctggtgctg ttgagttctg gactgataga ctcggtttct cccgtgactt cgtagaggac
     4981 gactttgccg gtgttgtacg tgacgacgtt accctgttca tctccgcagt tcaggaccag
     5041 gttgtgccag acaacactct ggcatgggta tgggttcgtg gtctggacga actgtacgct
     5101 gagtggtctg aggtcgtgtc taccaacttc cgtgatgcat ctggtccagc tatgaccgag
     5161 atcggtgaac agccctgggg tcgtgagttt gcactgcgtg atccagctgg taactgcgtg
     5221 catttcgtcg cagaagagca ggactaacaa ttgacacctt acgattattt agagagtatt
     5281 tattagtttt attgtatgta tacggatgtt ttattatcta tttatgccct tatattctgt
     5341 aactatccaa aagtcctatc ttatcaagcc agcaatctat gtccgcgaac gtcaactaaa
     5401 aataagcttt ttatgctctt ctctcttttt ttcccttcgg tataattata ccttgcatcc
     5461 acagattctc ctgccaaatt ttgcataatc ctttacaaca tggctatatg ggagcactta
     5521 gcgccctcca aaacccatat tgcctacgca tgtataggtg ttttttccac aatattttct
     5581 ctgtgctctc tttttattaa agagaagctc tatatcggag aagcttctgt ggccgttata
     5641 ttcggcctta tcgtgggacc acattgcctg aattggtttg ccccggaaga ttggggaaac
     5701 ttggatctga ttaccttagc tgcaggtacc actgagcgtc agaccccgta gaaaagatca
     5761 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5821 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5881 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5941 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6001 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6061 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6121 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6181 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6241 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6301 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6361 acgccagcaa cgcggccttt ttacggttcc tggccttttg c
//
