LOCUS       sequence-494832-        4976 bp    DNA     circular SYN 09-DEC-2025
DEFINITION  Yeast gene tagging with mAID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4976)
  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 4976)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 9, 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-mAID-mNeon-HIS3MX6
FEATURES             Location/Qualifiers
     source          1..4976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             63..266
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     terminator      1004..1191
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1059..1083)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1266..1609
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1610..2308
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      2312..2509
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2613..2632)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2614..2632)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2719..2736)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2890..3478)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2970..2989)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3649..4440,4441..4509))
                     /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     4272..4291
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4510..4614)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4682..4700
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4738..4760)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4860..4879
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4960..4976,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4960..4976,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acaaggagaa gagtgcttgt cctaaagatc cagccaaacc tccggccaag gcacaagttg
      121 tgggatggcc accggtgaga tcataccgga agaacgtgat ggtttcctgc caaaaatcaa
      181 gcggtggccc ggaggcggcg gcgttcgtga aggtatcaat ggacggagca ccgtacttga
      241 ggaaaatcga tttgaggatg tataaaggtt ctggtgtttc taagggtgaa gaagacaaca
      301 tggcttcttt gccagctact cacgaattgc acatcttcgg ttctatcaac ggtgttgact
      361 tcgacatggt tggtcaaggt actggtaacc caaacgacgg ttacgaagaa ttgaacttga
      421 agtctactaa gggtgacttg caattctctc catggatctt ggttccacac atcggttacg
      481 gtttccacca atacttgcca tacccagacg gtatgtctcc attccaagct gctatggttg
      541 acggttctgg ttaccaagtt cacagaacta tgcaattcga agacggtgct tctttgactg
      601 ttaactacag atacacttac gaaggttctc acatcaaggg tgaagctcaa gttaagggta
      661 ctggtttccc agctgacggt ccagttatga ctaactcttt gactgctgct gactggtgta
      721 gatctaagaa gacttaccca aacgacaaga ctatcatctc tactttcaag tggtcttaca
      781 ctactggtaa cggtaagaga tacagatcta ctgctagaac tacttacact ttcgctaagc
      841 caatggctgc taactacttg aagaaccaac caatgtacgt tttcagaaag actgaattga
      901 agcactctaa gactgaattg aacttcaagg aatggcaaaa ggctttcact gacgttatgg
      961 gtatggacga attgtacaag taaggcgcgc cacttctaaa taagcgaatt tcttatgatt
     1021 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     1081 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     1141 gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg gcagatccgc
     1201 tagggataac agggtaatat agatctgttt agcttgcctc gtccccgccg ggtcacccgg
     1261 ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg
     1321 catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat
     1381 ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct
     1441 gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt
     1501 agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta
     1561 aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca tggcagaacc
     1621 agcccaaaaa aagcaaaaac aaactgttca ggagcgcaag gcgtttatct cccgtatcac
     1681 taatgaaact aaaattcaaa tcgctatttc gctgaatggt ggttatattc aaataaaaga
     1741 ttcgattctt cctgcaaaga aggatgacga tgtagcttcc caagctactc agtcacaggt
     1801 catcgatatt cacacaggtg ttggcttttt ggatcatatg atccatgcgt tggcaaaaca
     1861 ctctggttgg tctcttattg ttgaatgtat tggtgacctg cacattgacg atcaccatac
     1921 taccgaagat tgcggtatcg cattagggca agcgttcaaa gaagcaatgg gtgctgtccg
     1981 tggtgtaaaa agattcggta ctgggttcgc accattggat gaggcgctat cacgtgccgt
     2041 agtcgattta tctagtagac catttgctgt aatcgacctt ggattgaaga gagagatgat
     2101 tggtgattta tccactgaaa tgattccaca ctttttggaa agtttcgcgg aggcggccag
     2161 aattactttg catgttgatt gtctgagagg tttcaacgat caccacagaa gtgagagtgc
     2221 gttcaaggct ttggctgttg ccataagaga agctatttct agcaatggca ccaatgacgt
     2281 tccctcaacc aaaggtgttt tgatgtgaag tactgacaat aaaaagattc ttgttttcaa
     2341 gaacttgtca tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag
     2401 cgtgatttat attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca
     2461 gaaagtaata tcatgcgtca atcgtatgtg aatgctggtc gctatactgc tgtcgattcg
     2521 atactaacgc cgccatccag tttaaacgag ctcgaattca tcgatgatat cagatccact
     2581 agtggcctat gcggccgcgg atctgccggt ctccctatag tgagtcgtat taatttcgat
     2641 aagccaggtt aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2701 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2761 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2821 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2881 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2941 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3001 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3061 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3121 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3181 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3241 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3301 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3361 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3421 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3481 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3541 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3601 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3661 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3721 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3781 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3841 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3901 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3961 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4021 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4081 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4141 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4201 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4261 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4321 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4381 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4441 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4501 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4561 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4621 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     4681 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     4741 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4801 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     4861 ggcatcagag cagattgtac tgagagtgca ccatatggac atattgtcgt tagaacgcgg
     4921 ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac actata
//
