LOCUS       sequence-494833-        4708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast gene tagging with mAID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4708)
  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 4708)
  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-mAID-mNeon-kITRP1
FEATURES             Location/Qualifiers
     source          1..4708
                     /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"
     primer_bind     complement(2345..2364)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2346..2364)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2451..2468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2622..3210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2702..2721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3381..4172,4173..4241))
                     /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     4004..4023
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4242..4346)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4414..4432
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4470..4492)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4592..4611
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4692..4708,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4692..4708,1)
                     /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 agatctaaag tggaacgatc attcactata tatatatcaa
     1261 tttatatata cgtatgtgta attgaagaaa gatacgtttt ttcctctatt gagaggcctg
     1321 ctggatgaat agctttacct tttctaaatc cttgatacca tcagtctcta ctcctccact
     1381 tacatcgaca ccgatcgcat ttggtaacat attaatggca acagaaacgt tatcaggatt
     1441 caatccacca gcgataatga attttatctc gggatgactt gcagaccaac tggaaattgc
     1501 actccaattc aatttctcac cagtgccacc ttcaccagaa tcgaacaacg tcagcacatt
     1561 gtctacgtgt tcatacaggt ccagtagtaa ttcacaatcc tgtgggaact ggaacctctt
     1621 aatgattgga attgaagatg ggatcaaaga tctgtattct ttaatatctt catctccatg
     1681 taattgtatc acatctagat tatattcgtg gtacagttga aggacatcat caacggactg
     1741 atttctaaac accccgacta gtttagtacc tttcacgttc tcttgttggt gaactgcagt
     1801 tgaaatacct ttcgcaacag atgagtcaat ggttcttttc ctaccgggaa cacaaatgat
     1861 acctaagtaa tcagcaccat catccacagc agtctttgca gcttcaacgg tttgcaaacc
     1921 acacacttta acgagcatca cagtagattt cgcaagaggg tagctcgctc agagtaccca
     1981 agtaaatgat tagtaaactg atgtttgata gttcaatttt tcaatgaaat aaccttatat
     2041 taaaattgat attactatta tacaaaaata aagaataaag gatttgagtt tatacataaa
     2101 ataccattat tatttgttca gtgagagata ccggggtata tgggatgtgt gtagtgatac
     2161 catgcaatca tgtatcaaac atggaagaag tgaaggggga gaaagaaact tgaaaaaaag
     2221 aaaaccattt gaaaagagga aaagtatcta attaagatct ttcacctctt agtttaaacg
     2281 agctcgaatt catcgatgat atcagatcca ctagtggcct atgcggccgc ggatctgccg
     2341 gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca ttaatgaatc
     2401 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2461 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2521 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2581 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2641 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2701 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2761 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2821 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2881 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2941 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3001 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3061 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3121 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3181 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3241 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3301 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3361 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3421 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3481 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3541 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3601 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3661 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3721 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3781 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3841 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3901 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3961 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4021 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4081 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4141 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4201 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4261 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4321 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     4381 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     4441 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     4501 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     4561 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     4621 caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc
     4681 atacacatac gatttaggtg acactata
//
