LOCUS       sequence-495239-        4932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast gene tagging with mIAA7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4932)
  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 4932)
  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-mIAA7-mNeon-natNT2
FEATURES             Location/Qualifiers
     source          1..4932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1004..1191
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     promoter        1266..1609
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1610..2182
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      2190..2377
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2569..2588)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2570..2588)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2675..2692)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2846..3434)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2926..2945)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3605..4396,4397..4465))
                     /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     4228..4247
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4466..4570)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4638..4656
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4694..4716)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4816..4835
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4916..4932,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4916..4932,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acggcttctc tgagaccgtg gacctgatgc tgaacctgca gtccaataag gagggctctg
      121 tggatctgaa gaacgtgagc gccgtgccta aggagaagac cacactgaag gacccatcca
      181 agccccctgc caaggcacag gtggtgggat ggccacccgt gcggaactac agaaagaata
      241 tgatgaccca gcagaagaca agctccggtt 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 tgggtaccac
     1621 tcttgacgac acggcttacc ggtaccgcac cagtgtcccg ggggacgccg aggccatcga
     1681 ggcactggat gggtccttca ccaccgacac cgtcttccgc gtcaccgcca ccggggacgg
     1741 cttcaccctg cgggaggtgc cggtggaccc gcccctgacc aaggtgttcc ccgacgacga
     1801 atcggacgac gaatcggacg acggggagga cggcgacccg gactcccgga cgttcgtcgc
     1861 gtacggggac gacggcgacc tggcgggctt cgtggtcgtc tcgtactccg gctggaaccg
     1921 ccggctgacc gtcgaggaca tcgaggtcgc cccggagcac cgggggcacg gggtcgggcg
     1981 cgcgttgatg gggctcgcga cggagttcgc ccgcgagcgg ggcgccgggc acctctggct
     2041 ggaggtcacc aacgtcaacg caccggcgat ccacgcgtac cggcggatgg ggttcaccct
     2101 ctgcggcctg gacaccgccc tgtacgacgg caccgcctcg gacggcgagc aggcgctcta
     2161 catgagcatg ccctgcccct aatctcgagg cgaatttctt atgatttatg atttttatta
     2221 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa
     2281 aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat
     2341 agcatgaggt cgctcttatt gaccacacct ctaccggcat gccgagcaaa tgcctgcaaa
     2401 tcgctcccca tttcacccaa ttgtagatat gctaactcca gcaatgagtt gatgaatctc
     2461 ggtgtgtatt ttatgtcctc agaggacaac acctgttgta atcgagctcg aattcatcga
     2521 tgatatcaga tccactagtg gcctatgcgg ccgcggatct gccggtctcc ctatagtgag
     2581 tcgtattaat ttcgataagc caggttaacc tgcattaatg aatcggccaa cgcgcgggga
     2641 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2701 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2761 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2821 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2881 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2941 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3001 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3061 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3121 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3181 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3241 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3301 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3361 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3421 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3481 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3541 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3601 acagttacca atgcttaatc agcgaggcac ctatctcagc gatctgtcta tttcgttcat
     3661 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3721 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3781 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3841 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3901 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3961 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4021 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4081 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4141 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4201 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4261 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4321 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4381 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4441 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4501 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4561 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     4621 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     4681 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     4741 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     4801 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atggacatat
     4861 tgtcgttaga acgcggctac aattaataca taaccttatg tatcatacac atacgattta
     4921 ggtgacacta ta
//
