LOCUS       sequence-392256-        6773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses OsTIR1F74A under the control of ADH1 promoter in budding 
            yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6773)
  AUTHORS   Ogawa Y, Nishimura K, Obara K, Kamura T
  TITLE     Development of AlissAID system targeting GFP or mCherry fusion 
            protein.
  JOURNAL   PLoS Genet. 2023 Jun 14;19(6):e1010731. doi: 
            10.1371/journal.pgen.1010731. eCollection 2023 Jun.
  PUBMED    37315088
REFERENCE   2  (bases 1 to 6773)
  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: "PLoS 
            Genet."; date: "2023-06-14"; volume: "19(6):e1010731. doi"; pages: "
            10.1371/journal.pgen.1010731. eCollection 2023 Jun"
COMMENT     Sequence Label: pRS306-OsTIR1F74A
FEATURES             Location/Qualifiers
     source          1..6773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2243..2261
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2289..2305
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2290..2306
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2613..3014
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     primer_bind     complement(4792..4811)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4793..4811)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4818..4835)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4818..4834)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4827..4849)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4975..5430
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5062..5081)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5272..5293
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5561..6364)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(6365..6580)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6604..6623)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6723..6745
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctc gaaattaacc ctcactaaag ggaacaaaag ctggtaccgg
     2281 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcgggt gtacaatatg
     2341 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
     2401 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
     2461 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
     2521 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
     2581 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
     2641 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac
     2701 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
     2761 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc
     2821 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt
     2881 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
     2941 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
     3001 gcatacaatc aactatctca tatacaacta gtatgactta ttttcctgaa gaagttgtag
     3061 aacacatatt ttccttttta cctgctcaaa gagatagaaa caccgtctcc ttggtctgta
     3121 aggtttggta cgaaatcgaa agattgtcta gaagaggtgt ttttgttggt aattgttacg
     3181 cagttagagc tggtagagtt gctgcaagat tcccaaacgt tagagcattg actgttaaag
     3241 gtaaacctca tgcagctgat ttcaatttgg ttccacctga ttggggtggt tatgctggtc
     3301 catggattga agctgcagct agaggttgtc acggtttgga agaattgaga atgaagagaa
     3361 tggttgtttc tgatgaatca ttggaattgt tagcaagatc ttttcctaga ttcagagctt
     3421 tggttttgat ctcatgtgaa ggtttttcca ccgatggttt agcagctgtt gcttcacatt
     3481 gtaagttgtt gagagaattg gatttgcaag aaaacgaagt tgaagataga ggtccaagat
     3541 ggttatcttg ttttcctgat tcctgtacaa gtttggtttc tttgaacttc gcatgtatca
     3601 agggtgaagt taacgctggt agtttggaaa gattagtttc tagatcacca aatttgagat
     3661 ctttgagatt gaacagatcc gttagtgttg ataccttggc aaagatcttg ttgagaactc
     3721 ctaatttgga agatttgggt actggtaact tgacagatga tttccaaaca gaatcatact
     3781 tcaagttgac ctccgctttg gaaaagtgta agatgttgag atctttgtca ggtttttggg
     3841 atgcaagtcc agtttgtttg tcttttatat atcctttgtg tgctcaattg actggtttga
     3901 atttgtctta cgcaccaact ttggatgcta gtgatttgac aaagatgatc tctagatgtg
     3961 ttaagttgca aagattgtgg gttttggatt gtatctcaga taaaggtttg caagttgttg
     4021 cttcttcttg taaggatttg caagaattga gagtttttcc aagtgatttc tatgttgcag
     4081 gttactctgc tgttacagaa gaaggtttag ttgcagtttc attgggttgt cctaagttga
     4141 actccttgtt gtacttctgt catcaaatga ccaacgcagc tttggttact gttgctaaaa
     4201 attgtccaaa cttcactaga ttcagattgt gtatcttgga accaggtaaa cctgatgttg
     4261 ttacatcaca acctttagat gaaggttttg gtgcaatcgt tagagagtgt aagggtttgc
     4321 aaagattgtc catcagtggt ttgttgacag ataaggtttt tatgtacatc ggtaaatacg
     4381 ctaagcaatt ggaaatgttg tcaatcgcat tcgctggtga ctccgataag ggtatgatgc
     4441 atgttatgaa cggttgtaag aacttaagaa agttggaaat cagagattct ccatttggtg
     4501 acgcagcttt gttgggtaat ttcgcaagat atgaaacaat gagatctttg tggatgtcca
     4561 gttgtaacgt taccttgaaa ggttgtcaag ttttagcttc aaagatgcca atgttgaacg
     4621 ttgaagttat taacgaaaga gatggttcca atgaaatgga agaaaaccat ggtgacttgc
     4681 ctaaagttga aaagttatac gtctacagaa caaccgcagg tgccagagat gatgctccta
     4741 acttcgtcaa gattttatga gcggccgcca ccgcggtgga gctccaattc gccctatagt
     4801 gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     4861 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     4921 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc
     4981 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5041 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5101 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5161 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5221 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5281 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5341 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5401 attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     5461 gtgcggtatt tcacaccgca tagggtaata actgatataa ttaaattgaa gctctaattt
     5521 gtgagtttag tatacatgca tttacttata atacagtttt ttagttttgc tggccgcatc
     5581 ttctcaaata tgcttcccag cctgcttttc tgtaacgttc accctctacc ttagcatccc
     5641 ttccctttgc aaatagtcct cttccaacaa taataatgtc agatcctgta gagaccacat
     5701 catccacggt tctatactgt tgacccaatg cgtctccctt gtcatctaaa cccacaccgg
     5761 gtgtcataat caaccaatcg taaccttcat ctcttccacc catgtctctt tgagcaataa
     5821 agccgataac aaaatctttg tcgctcttcg caatgtcaac agtaccctta gtatattctc
     5881 cagtagatag ggagcccttg catgacaatt ctgctaacat caaaaggcct ctaggttcct
     5941 ttgttacttc ttctgccgcc tgcttcaaac cgctaacaat acctgggccc accacaccgt
     6001 gtgcattcgt aatgtctgcc cattctgcta ttctgtatac acccgcagag tactgcaatt
     6061 tgactgtatt accaatgtca gcaaattttc tgtcttcgaa gagtaaaaaa ttgtacttgg
     6121 cggataatgc ctttagcggc ttaactgtgc cctccatgga aaaatcagtc aagatatcca
     6181 catgtgtttt tagtaaacaa attttgggac ctaatgcttc aactaactcc agtaattcct
     6241 tggtggtacg aacatccaat gaagcacaca agtttgtttg cttttcgtgc atgatattaa
     6301 atagcttggc agcaacagga ctaggatgag tagcagcacg ttccttatat gtagctttcg
     6361 acatgattta tcttcgtttc ctgcaggttt ttgttctgtg cagttgggtt aagaatactg
     6421 ggcaatttca tgtttcttca acactacata tgcgtatata taccaatcta agtctgtgct
     6481 ccttccttcg ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc
     6541 gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa aaggtggtat ggtgcactct
     6601 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     6661 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     6721 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cga
//
