LOCUS       sequence-494854-        9264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of OsTIR1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9264)
  AUTHORS   Hubbe P, Sharma C, Pajonk O, Peters N, Guschtschin-Schmidt N, 
            Friemel N, Schuck S
  TITLE     An optimised system for rapid auxin-inducible protein degradation in
            budding yeast
  JOURNAL   bioRxiv 2025.09.24.678194
REFERENCE   2  (bases 1 to 9264)
  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: "bioRxiv 
            2025.09.24.678194"
COMMENT     Sequence Label: pNH605-Padh-OsTIR1(F74A)
FEATURES             Location/Qualifiers
     source          1..9264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        276..683
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     promoter        2509..3213
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     primer_bind     complement(4980..4996)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4981..4997)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4999..5246
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5015..5033)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5811..5830)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5812..5830)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5837..5854)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5837..5853)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5846..5868)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5994..6449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6081..6100)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6291..6312
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6805..6824)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6924..6946
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6984..7002)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7087..7191
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7192..7260,7261..8052)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7410..7429)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8223..8811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8712..8731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8965..8982
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9099..9120
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9135..9140,9141..9158,9159..9165)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9173..9189
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     9178..9200
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9197..9213
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9197..9213
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        9234..9252
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggccggccat ttaaatgttt aaacggccac gctggccgag gagaacttct agtatatcta
       61 catacctaat attattgcct tattaaaaat ggaatcccaa cgattacatc aaaatccaca
      121 ttctcttcaa aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa aacgttatat
      181 ttataggata attatactct atttctcaac aagtaattgg ttgtttggcc gagcggtcta
      241 aggcgcctga ttcaagaaat atcttgaccg cagttaactg tgggaatact caggtatcgt
      301 aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca taacgagaac
      361 acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt gttaatttca
      421 gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga
      481 gagcgccgga accggctttt catatagaat agagaagcgt tcatgactaa atgcttgcat
      541 cacaatactt gaagttgaca atattattta aggacctatt gttttttcca ataggtggtt
      601 agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata tatatatata
      661 tatttcaagg atataccatt ctaatgtctg cccctaagtt accaatagat agcggtttaa
      721 tataaggttt caacaagaaa tattttgtga aaatcaaaag ttaatagttt cactgtcgaa
      781 aatatgaaag tttaagtttt cagttcatcc cattgtagtt atcaaatttc tgtgtttccc
      841 gtatatgtgt gctctgtatt catcgctatc ttgaaactgg ttatgatcgt tgcgcatgtg
      901 acctgtatcg ctcaaaagag cccggcaccg gtaccgggca aaagccccta gcaagctaaa
      961 atgattcaac tttcgtagta atttgttcaa ccggtcacag taaagtcaat gactaacttt
     1021 tgaatcgagg atgtatttcc aattgaaagc aatgtatata taaaccattg tgttgaagct
     1081 tgtcttacag ttttcttgta tcctcccatt gttcatatag tacttaaagt accataatcc
     1141 ttaaagaaca aacttgaaag agaatacact atacacaact aataaataaa gaataatcat
     1201 ggctgtgacc aagacaattg tagttctacc aggtgaccat gttggtcaag aaatcactga
     1261 agaggccatt aaagttttaa atgctattca ggaatgtcgt ccagacaagg tcaatttcaa
     1321 gtttgatcat catttgatcg gtggtgctgc aattgatgcc actggtgttc cattaccaga
     1381 cgaagctttg gaggcctcta agaaagctga tgccgtgctt ctgggtgctg ttggtggtcc
     1441 aaaatggggt actggcgctg tcagaccaga acaaggtctt ttgaaaatcc gtaaggagtt
     1501 gcaattgtat gctaatctaa gaccatgtaa ttttgcatct gattccttac tagatctatc
     1561 gccattgaag cctgaaattg caagaggtac agatttcgtc gttgttagag aactagtggg
     1621 tggtatttat ttcggggaga gaaaagaaga tgaaggtgat ggtgtcgcct gggatagcga
     1681 aaagtattct gtgcccgaag ttcaaagaat cactagaatg gctgctttca tggctctgca
     1741 acacaaccca cctttaccta tttggtctct agataaagcc aacgttctag catcatcaag
     1801 attatggaga aaaacagtgg aagagacgat taagaatgaa ttcccacaat tgactgttca
     1861 acatcaattg attgattccg ctgctatgat cttggttaag aacccaactc atctaaatgg
     1921 tattattatc acaaacaata tgtttggtga tattatctcc gatgaagcct ccgtgatccc
     1981 aggttcgtta ggattacttc catctgcctc cttggcatct cttccagata agaatacggc
     2041 atttggttta tatgagccat gtcatggctc tgctccagac ttaccaaagg gaaaggttaa
     2101 ccccgtagca acaatcctat ctgctgctat gatgttgaag ttgtctctgg acctgttcga
     2161 agaaggtgaa attatagaac aagcagtcaa gaaagtgttg gattctggta tcagaacagc
     2221 ggatctaaag ggtaccaact ctactactga ggtcggtgat gcagtagcga aagctgtcag
     2281 ggaactatta gcttagactt agtagatgta cagttgcata cataagtctg aacagaaaga
     2341 agtaaatatc atatctagtt cggtttagaa atcagtgact atattggttt tgaccagatc
     2401 gcaatccttg caagcattta gtaatgacta tcaaaccggt gaatcatctg ataaggctat
     2461 attagcttgt gcattcgcat gtatcgggaa acgaacggta ccgggcccat ccttttgttg
     2521 tttccgggtg tacaatatgg acttcctctt ttctggcaac caaacccata catcgggatt
     2581 cctataatac cttcgttggt ctccctaaca tgtaggtggc ggaggggaga tatacaatag
     2641 aacagatacc agacaagaca taatgggcta aacaagacta caccaattac actgcctcat
     2701 tgatggtggt acataacgaa ctaatactgt agccctagac ttgatagcca tcatcatatc
     2761 gaagtttcac tacccttttt ccatttgcca tctattgaag taataatagg cgcatgcaac
     2821 ttcttttctt tttttttctt ttctctctcc cccgttgttg tctcaccata tccgcaatga
     2881 caaaaaaatg atggaagaca ctaaaggaaa aaattaacga caaagacagc accaacagat
     2941 gtcgttgttc cagagctgat gaggggtatc tcgaagcaca cgaaactttt tccttccttc
     3001 attcacgcac actactctct aatgagcaac ggtatacggc cttccttcca gttacttgaa
     3061 tttgaaataa aaaaaagttt gctgtcttgc tatcaagtat aaatagacct gcaattatta
     3121 atcttttgtt tcctcgtcat tgttctcgtt ccctttcttc cttgtttctt tttctgcaca
     3181 atatttcaag ctataccaag catacaatca actccaagct ggccgctcta gaactagtat
     3241 gacttatttt cctgaagaag ttgtagaaca catattttcc tttttacctg ctcaaagaga
     3301 tagaaacacc gtctccttgg tctgtaaggt ttggtacgaa atcgaaagat tgtctagaag
     3361 aggtgttttt gttggtaatt gttacgcagt tagagctggt agagttgctg caagattccc
     3421 aaacgttaga gcattgactg ttaaaggtaa acctcatgcg gctgatttca atttggttcc
     3481 acctgattgg ggtggttatg ctggtccatg gattgaagct gcagctagag gttgtcacgg
     3541 tttggaagaa ttgagaatga agagaatggt tgtttctgat gaatcattgg aattgttagc
     3601 aagatctttt cctagattca gagctttggt tttgatctca tgtgaaggtt tttccaccga
     3661 tggtttagca gctgttgctt cacattgtaa gttgttgaga gaattggatt tgcaagaaaa
     3721 cgaagttgaa gatagaggtc caagatggtt atcttgtttt cctgattcct gtacaagttt
     3781 ggtttctttg aacttcgcat gtatcaaggg tgaagttaac gctggtagtt tggaaagatt
     3841 agtttctaga tcaccaaatt tgagatcttt gagattgaac agatccgtta gtgttgatac
     3901 cttggcaaag atcttgttga gaactcctaa tttggaagat ttgggtactg gtaacttgac
     3961 agatgatttc caaacagaat catacttcaa gttgacctcc gctttggaaa agtgtaagat
     4021 gttgagatct ttgtcaggtt tttgggatgc aagtccagtt tgtttgtctt ttatatatcc
     4081 tttgtgtgct caattgactg gtttgaattt gtcttacgca ccaactttgg atgctagtga
     4141 tttgacaaag atgatctcta gatgtgttaa gttgcaaaga ttgtgggttt tggattgtat
     4201 ctcagataaa ggtttgcaag ttgttgcttc ttcttgtaag gatttgcaag aattgagagt
     4261 ttttccaagt gatttctatg ttgcaggtta ctctgctgtt acagaagaag gtttagttgc
     4321 agtttcattg ggttgtccta agttgaactc cttgttgtac ttctgtcatc aaatgaccaa
     4381 cgcagctttg gttactgttg ctaaaaattg tccaaacttc actagattca gattgtgtat
     4441 cttggaacca ggtaaacctg atgttgttac atcacaacct ttagatgaag gttttggtgc
     4501 aatcgttaga gagtgtaagg gtttgcaaag attgtccatc agtggtttgt tgacagataa
     4561 ggtttttatg tacatcggta aatacgctaa gcaattggaa atgttgtcaa tcgcattcgc
     4621 tggtgactcc gataagggta tgatgcatgt tatgaacggt tgtaagaact taagaaagtt
     4681 ggaaatcaga gattctccat ttggtgacgc agctttgttg ggtaatttcg caagatatga
     4741 aacaatgaga tctttgtgga tgtccagttg taacgttacc ttgaaaggtt gtcaagtttt
     4801 agcttcaaag atgccaatgt tgaacgttga agttattaac gaaagagatg gttccaatga
     4861 aatggaagaa aaccatggtg acttgcctaa agttgaaaag ttatacgtct acagaacaac
     4921 cgcaggtgcc agagatgatg ctcctaactt cgtcaagatt ttatgagcgg caagcttatc
     4981 gataccgtcg acctcgagtc atgtaattag ttatgtcacg cttacattca cgccctcccc
     5041 ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt
     5101 atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     5161 ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt
     5221 tgggacgctc gaaggcttta atttgctggc caccgaagtc ggtgatgctg tcgccgaaga
     5281 agttaagaaa atccttgctt aaaaagattc tcttttttta tgatatttgt acataaactt
     5341 tataaatgaa attcataata gaaacgacac gaaattacaa aatggaatat gttcataggg
     5401 tagacgaaac tatatacgca atctacatac atttatcaag aaggagaaaa aggaggatgt
     5461 aaaggaatac aggtaagcaa attgatacta atggctcaac gtgataagga aaaagaattg
     5521 cactttaaca ttaatattga caaggaggag ggcaccacac aaaaagttag gtgtaacaga
     5581 aaatcatgaa actatgattc ctaatttata tattggagga ttttctctaa aaaaaaaaaa
     5641 tacaacaaat aaaaaacact caatgacctg accatttgat ggagtttaag tcaatacctc
     5701 cttgaaccat ttcccataat ggtgaaagtt ccctcaagaa ttttactctg tcagaaacgg
     5761 ccttaacgac gtaggccggc catttaaatg tttaaacggc cacgctggcc ccctatagtg
     5821 agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     5881 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     5941 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc
     6001 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     6061 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     6121 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     6181 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     6241 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     6301 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     6361 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     6421 ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     6481 tgcggtattt cacaccgcat agggtaataa ctgatataat taaattgaag ctctaatttg
     6541 tgagtttagt attaggatct ctacccttgg cgaaaagtcc tctgccaaca atgatgatat
     6601 ctgatccacc acttacaact tcgtcgacgg ttctgtactg ctgacccaat atgcgtatat
     6661 ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag attaccgaat
     6721 caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga
     6781 aaaggtggta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     6841 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     6901 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     6961 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     7021 gataataatg gtttcttaga cgtcggcgcg ccgaggacgt caggtggcac ttttcgggga
     7081 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     7141 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     7201 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     7261 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     7321 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     7381 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     7441 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     7501 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     7561 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     7621 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     7681 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     7741 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     7801 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     7861 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     7921 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     7981 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     8041 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     8101 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     8161 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     8221 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     8281 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     8341 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     8401 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     8461 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     8521 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     8581 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     8641 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     8701 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     8761 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     8821 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     8881 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     8941 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     9001 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     9061 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     9121 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     9181 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct cggaattaac
     9241 cctcactaaa gggaacaaaa gctg
//
