LOCUS       sequence-494871-        4904 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast gene tagging with AID*.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4904)
  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 4904)
  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: pFA6a-AID*-mNeon-HIS3MX6
FEATURES             Location/Qualifiers
     source          1..4904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             63..194
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Morawska and Ulrich, 
                     2013)"
                     /label=AID*
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="PKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEAAAFVK"
     terminator      932..1119
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(987..1011)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1194..1537
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1538..2236
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      2240..2437
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2541..2560)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2542..2560)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2647..2664)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2818..3406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2898..2917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3577..4368,4369..4437))
                     /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     4200..4219
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4438..4542)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4610..4628
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4666..4688)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4788..4807
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4888..4904,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4888..4904,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 accctaaaga tccagccaaa cctccggcca aggcacaagt tgtgggatgg ccaccggtga
      121 gatcataccg gaagaacgtg atggtttcct gccaaaaatc aagcggtggc ccggaggcgg
      181 cggcgttcgt gaagggttct ggtgtttcta agggtgaaga agacaacatg gcttctttgc
      241 cagctactca cgaattgcac atcttcggtt ctatcaacgg tgttgacttc gacatggttg
      301 gtcaaggtac tggtaaccca aacgacggtt acgaagaatt gaacttgaag tctactaagg
      361 gtgacttgca attctctcca tggatcttgg ttccacacat cggttacggt ttccaccaat
      421 acttgccata cccagacggt atgtctccat tccaagctgc tatggttgac ggttctggtt
      481 accaagttca cagaactatg caattcgaag acggtgcttc tttgactgtt aactacagat
      541 acacttacga aggttctcac atcaagggtg aagctcaagt taagggtact ggtttcccag
      601 ctgacggtcc agttatgact aactctttga ctgctgctga ctggtgtaga tctaagaaga
      661 cttacccaaa cgacaagact atcatctcta ctttcaagtg gtcttacact actggtaacg
      721 gtaagagata cagatctact gctagaacta cttacacttt cgctaagcca atggctgcta
      781 actacttgaa gaaccaacca atgtacgttt tcagaaagac tgaattgaag cactctaaga
      841 ctgaattgaa cttcaaggaa tggcaaaagg ctttcactga cgttatgggt atggacgaat
      901 tgtacaagta aggcgcgcca cttctaaata agcgaatttc ttatgattta tgatttttat
      961 tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt
     1021 aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt
     1081 atagtatgag gtcgctctta ttgaccacac ctctaccggc agatccgcta gggataacag
     1141 ggtaatatag atctgtttag cttgcctcgt ccccgccggg tcacccggcc agcgacatgg
     1201 aggcccagaa taccctcctt gacagtcttg acgtgcgcag ctcaggggca tgatgtgact
     1261 gtcgcccgta catttagccc atacatcccc atgtataatc atttgcatcc atacattttg
     1321 atggccgcac ggcgcgaagc aaaaattacg gctcctcgct gcagacctgc gagcagggaa
     1381 acgcacccct cacagacgcg ttgaattgtc cccacgccgc gcccctgtag agaaatataa
     1441 aaggttagga tttgccactg aggttcttct ttcatatact tccttttaaa atcttgctag
     1501 gatacagttc tcacatcaca tccgaacata aacaaccatg gcagaaccag cccaaaaaaa
     1561 gcaaaaacaa actgttcagg agcgcaaggc gtttatctcc cgtatcacta atgaaactaa
     1621 aattcaaatc gctatttcgc tgaatggtgg ttatattcaa ataaaagatt cgattcttcc
     1681 tgcaaagaag gatgacgatg tagcttccca agctactcag tcacaggtca tcgatattca
     1741 cacaggtgtt ggctttttgg atcatatgat ccatgcgttg gcaaaacact ctggttggtc
     1801 tcttattgtt gaatgtattg gtgacctgca cattgacgat caccatacta ccgaagattg
     1861 cggtatcgca ttagggcaag cgttcaaaga agcaatgggt gctgtccgtg gtgtaaaaag
     1921 attcggtact gggttcgcac cattggatga ggcgctatca cgtgccgtag tcgatttatc
     1981 tagtagacca tttgctgtaa tcgaccttgg attgaagaga gagatgattg gtgatttatc
     2041 cactgaaatg attccacact ttttggaaag tttcgcggag gcggccagaa ttactttgca
     2101 tgttgattgt ctgagaggtt tcaacgatca ccacagaagt gagagtgcgt tcaaggcttt
     2161 ggctgttgcc ataagagaag ctatttctag caatggcacc aatgacgttc cctcaaccaa
     2221 aggtgttttg atgtgaagta ctgacaataa aaagattctt gttttcaaga acttgtcatt
     2281 tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat
     2341 tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc
     2401 atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg
     2461 ccatccagtt taaacgagct cgaattcatc gatgatatca gatccactag tggcctatgc
     2521 ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa gccaggttaa
     2581 cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2641 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2701 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2761 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2821 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2881 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2941 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3001 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3061 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3121 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3181 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3241 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3301 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3361 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3421 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3481 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3541 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3601 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3661 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3721 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3781 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3841 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3901 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3961 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4021 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4081 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4141 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4201 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4261 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4321 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4381 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4441 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4501 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4561 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     4621 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     4681 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     4741 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     4801 gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct acaattaata
     4861 cataacctta tgtatcatac acatacgatt taggtgacac tata
//
