LOCUS       sequence-494911-        4596 bp    DNA     circular SYN 09-DEC-2025
DEFINITION  Yeast gene tagging with mIAA7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4596)
  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 4596)
  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-mIAA7-3xFLAG-kanNP1
FEATURES             Location/Qualifiers
     source          1..4596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             265..330
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      354..541
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(409..433)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        595..1114
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     CDS             1115..1924
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1182..1201)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1930..2127
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2231..2250)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2232..2250)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2337..2354)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2508..3096)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2588..2607)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3267..4058,4059..4127))
                     /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     3890..3909
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4128..4232)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4300..4318
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4356..4378)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4478..4497
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4578..4596
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4578..4595
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 ggcttctctg agaccgtgga cctgatgctg aacctgcagt ccaataagga gggctctgtg
      121 gatctgaaga acgtgagcgc cgtgcctaag gagaagacca cactgaagga cccatccaag
      181 ccccctgcca aggcacaggt ggtgggatgg ccacccgtgc ggaactacag aaagaatatg
      241 atgacccagc agaagacaag ctccgactac aaagaccatg acggtgatta taaagatcat
      301 gacatcgatt acaaggatga cgatgacaag tagggcgcgc cacttctaaa taagcgaatt
      361 tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat
      421 tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt
      481 aggtcaggtt gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg
      541 gcagatccgc tagggataac agggtaatat agatctgttt agcttgcctc gtcccctacg
      601 ttagtgtgag cgggatttaa actgtgagga ccttaataca ttcagacact tctgcggtat
      661 caccctactt attcccttcg agattatatc taggaaccca tcaggttggt ggaagattac
      721 ccgttctaag acttttcagc ttcctctatt gatgttacac ctggacaccc cttttctggc
      781 atccagtttt taatcttcag tggcatgtga gattctccga aattaattaa agcaatcaca
      841 caattctctc ggataccacc tcggttgaaa ctgacaggtg gtttgttacg catgctaatg
      901 caaaggagcc tatatacctt tggctcggct gctgtaacag ggaatataaa gggcagcata
      961 atttaggagt ttagtgaact tgcaacattt actattttcc cttcttacgt aaatattttt
     1021 ctttttaatt ctaaatcaat ctttttcaat tttttgtttg tattcttttc ttgcttaaat
     1081 ctataactac aaaaaacaca tacataaact aaaaatgggt aaggaaaaga ctcacgtttc
     1141 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     1201 taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga
     1261 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     1321 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     1381 tgatgatgca tggttactca ccactgcgat ccccggcaaa acagcattcc aggtattaga
     1441 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     1501 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     1561 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     1621 tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga
     1681 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     1741 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     1801 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     1861 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     1921 ctaatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt tgtatagttt
     1981 ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat tttttttcgc
     2041 ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat
     2101 cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg ccatccagtt
     2161 taaacgagct cgaattcatc gatgatatca gatccactag tggcctatgc ggccgcggat
     2221 ctgccggtct ccctatagtg agtcgtatta atttcgataa gccaggttaa cctgcattaa
     2281 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2341 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2401 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2461 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2521 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2581 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2641 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2701 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2761 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2821 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2881 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2941 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3001 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3061 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3121 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3181 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3241 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3301 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3361 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3421 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3481 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3541 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3601 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3661 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3721 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3781 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3841 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3901 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3961 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4021 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4081 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4141 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4201 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4261 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     4321 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     4381 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     4441 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     4501 agagtgcacc atatggacat attgtcgtta gaacgcggct acaattaata cataacctta
     4561 tgtatcatac acatacgatt taggtgacac tataga
//
