LOCUS       sequence-495241-        5244 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 5244)
  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 5244)
  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-kanNP1
FEATURES             Location/Qualifiers
     source          1..5244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1002..1189
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1057..1081)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1243..1762
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     CDS             1763..2572
                     /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(1830..1849)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2578..2775
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2879..2898)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2880..2898)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2985..3002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3156..3744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3236..3255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3915..4706,4707..4775))
                     /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     4538..4557
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4776..4880)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4948..4966
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5004..5026)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5126..5145
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5226..5244
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5226..5243
                     /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 ctccggttct ggtgtttcta agggtgaaga agacaacatg
      301 gcttctttgc cagctactca cgaattgcac atcttcggtt ctatcaacgg tgttgacttc
      361 gacatggttg gtcaaggtac tggtaaccca aacgacggtt acgaagaatt gaacttgaag
      421 tctactaagg gtgacttgca attctctcca tggatcttgg ttccacacat cggttacggt
      481 ttccaccaat acttgccata cccagacggt atgtctccat tccaagctgc tatggttgac
      541 ggttctggtt accaagttca cagaactatg caattcgaag acggtgcttc tttgactgtt
      601 aactacagat acacttacga aggttctcac atcaagggtg aagctcaagt taagggtact
      661 ggtttcccag ctgacggtcc agttatgact aactctttga ctgctgctga ctggtgtaga
      721 tctaagaaga cttacccaaa cgacaagact atcatctcta ctttcaagtg gtcttacact
      781 actggtaacg gtaagagata cagatctact gctagaacta cttacacttt cgctaagcca
      841 atggctgcta actacttgaa gaaccaacca atgtacgttt tcagaaagac tgaattgaag
      901 cactctaaga ctgaattgaa cttcaaggaa tggcaaaagg ctttcactga cgttatgggt
      961 atggacgaat tgtacaagta aggcgcgcca cttctaaata agcgaatttc ttatgattta
     1021 tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact
     1081 cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc
     1141 tttctcaggt atagtatgag gtcgctctta ttgaccacac ctctaccggc agatccgcta
     1201 gggataacag ggtaatatag atctgtttag cttgcctcgt cccctacgtt agtgtgagcg
     1261 ggatttaaac tgtgaggacc ttaatacatt cagacacttc tgcggtatca ccctacttat
     1321 tcccttcgag attatatcta ggaacccatc aggttggtgg aagattaccc gttctaagac
     1381 ttttcagctt cctctattga tgttacacct ggacacccct tttctggcat ccagttttta
     1441 atcttcagtg gcatgtgaga ttctccgaaa ttaattaaag caatcacaca attctctcgg
     1501 ataccacctc ggttgaaact gacaggtggt ttgttacgca tgctaatgca aaggagccta
     1561 tatacctttg gctcggctgc tgtaacaggg aatataaagg gcagcataat ttaggagttt
     1621 agtgaacttg caacatttac tattttccct tcttacgtaa atatttttct ttttaattct
     1681 aaatcaatct ttttcaattt tttgtttgta ttcttttctt gcttaaatct ataactacaa
     1741 aaaacacata cataaactaa aaatgggtaa ggaaaagact cacgtttcga ggccgcgatt
     1801 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     1861 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
     1921 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     1981 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     2041 gttactcacc actgcgatcc ccggcaaaac agcattccag gtattagaag aatatcctga
     2101 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     2161 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     2221 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     2281 tgaacaagtc tggaaagaaa tgcataagct tttgccattc tcaccggatt cagtcgtcac
     2341 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     2401 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     2461 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     2521 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagtact
     2581 gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta
     2641 gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc
     2701 tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg
     2761 ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccagttta aacgagctcg
     2821 aattcatcga tgatatcaga tccactagtg gcctatgcgg ccgcggatct gccggtctcc
     2881 ctatagtgag tcgtattaat ttcgataagc caggttaacc tgcattaatg aatcggccaa
     2941 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3001 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3061 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3121 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3181 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3241 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3301 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3361 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3421 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3481 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3541 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3601 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3661 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3721 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3781 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3841 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3901 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3961 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4021 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4081 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4141 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4201 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4261 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4321 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4381 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4441 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4501 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4561 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4621 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4681 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4741 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4801 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4861 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     4921 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     4981 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     5041 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     5101 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     5161 atggacatat tgtcgttaga acgcggctac aattaataca taaccttatg tatcatacac
     5221 atacgattta ggtgacacta taga
//
