LOCUS       sequence-494869-        4330 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 4330)
  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 4330)
  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-mIAA7-3xFLAG-HIS3MX6
FEATURES             Location/Qualifiers
     source          1..4330
                     /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        616..959
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             960..1658
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1664..1861
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1965..1984)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1966..1984)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2071..2088)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2242..2830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2322..2341)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3001..3792,3793..3861))
                     /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     3624..3643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3862..3966)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4034..4052
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4090..4112)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4212..4231
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4312..4330
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4312..4329
                     /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 gtccccgccg
      601 ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc
      661 agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa
      721 tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg
      781 ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc
      841 gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata
      901 cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca
      961 tggcagaacc agcccaaaaa aagcaaaaac aaactgttca ggagcgcaag gcgtttatct
     1021 cccgtatcac taatgaaact aaaattcaaa tcgctatttc gctgaatggt ggttatattc
     1081 aaataaaaga ttcgattctt cctgcaaaga aggatgacga tgtagcttcc caagctactc
     1141 agtcacaggt catcgatatt cacacaggtg ttggcttttt ggatcatatg atccatgcgt
     1201 tggcaaaaca ctctggttgg tctcttattg ttgaatgtat tggtgacctg cacattgacg
     1261 atcaccatac taccgaagat tgcggtatcg cattagggca agcgttcaaa gaagcaatgg
     1321 gtgctgtccg tggtgtaaaa agattcggta ctgggttcgc accattggat gaggcgctat
     1381 cacgtgccgt agtcgattta tctagtagac catttgctgt aatcgacctt ggattgaaga
     1441 gagagatgat tggtgattta tccactgaaa tgattccaca ctttttggaa agtttcgcgg
     1501 aggcggccag aattactttg catgttgatt gtctgagagg tttcaacgat caccacagaa
     1561 gtgagagtgc gttcaaggct ttggctgttg ccataagaga agctatttct agcaatggca
     1621 ccaatgacgt tccctcaacc aaaggtgttt tgatgtgatc agtactgaca ataaaaagat
     1681 tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg ttctatttta
     1741 atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc cagatgcgaa
     1801 gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg tcgctatact
     1861 gctgtcgatt cgatactaac gccgccatcc agtttaaacg agctcgaatt catcgatgat
     1921 atcagatcca ctagtggcct atgcggccgc ggatctgccg gtctccctat agtgagtcgt
     1981 attaatttcg ataagccagg ttaacctgca ttaatgaatc ggccaacgcg cggggagagg
     2041 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2101 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2161 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2221 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2281 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2341 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2401 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2461 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2521 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2581 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2641 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2701 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2761 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2821 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2881 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2941 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3001 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3061 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3961 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4021 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     4081 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     4141 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     4201 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgg acatattgtc
     4261 gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg
     4321 acactataga
//
