LOCUS       sequence-495247-        4302 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 4302)
  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 4302)
  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-ALFA-HIS3MX6
FEATURES             Location/Qualifiers
     source          1..4302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             265..303
                     /codon_start=1
                     /product="rationally designed epitope tag that forms a 
                     stable alpha-helix (Gotzke et al., 2019)"
                     /label=ALFA-tag
                     /note="It may be beneficial to flank the ALFA-tag sequence 
                     with proline residues to preserve the helical structure."
                     /translation="SRLEEELRRRLTE"
     terminator      326..513
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(381..405)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        588..931
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             932..1630
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1636..1833
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1937..1956)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1938..1956)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2043..2060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2214..2802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2294..2313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2973..3764,3765..3833))
                     /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     3596..3615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3834..3938)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4006..4024
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4062..4084)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4184..4203
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4284..4302
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4284..4301
                     /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 ctcctcaaga ttagaagagg agctgaggag aagattgacc
      301 gagtaagcgc gccacttcta aataagcgaa tttcttatga tttatgattt ttattattaa
      361 ataagttata aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg
      421 aaaattctta ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagta
      481 tgaggtcgct cttattgacc acacctctac cggcagatcc gctagggata acagggtaat
      541 atagatctgt ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc
      601 agaataccct ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc
      661 cgtacattta gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc
      721 gcacggcgcg aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc
      781 ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt
      841 aggatttgcc actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca
      901 gttctcacat cacatccgaa cataaacaac catggcagaa ccagcccaaa aaaagcaaaa
      961 acaaactgtt caggagcgca aggcgtttat ctcccgtatc actaatgaaa ctaaaattca
     1021 aatcgctatt tcgctgaatg gtggttatat tcaaataaaa gattcgattc ttcctgcaaa
     1081 gaaggatgac gatgtagctt cccaagctac tcagtcacag gtcatcgata ttcacacagg
     1141 tgttggcttt ttggatcata tgatccatgc gttggcaaaa cactctggtt ggtctcttat
     1201 tgttgaatgt attggtgacc tgcacattga cgatcaccat actaccgaag attgcggtat
     1261 cgcattaggg caagcgttca aagaagcaat gggtgctgtc cgtggtgtaa aaagattcgg
     1321 tactgggttc gcaccattgg atgaggcgct atcacgtgcc gtagtcgatt tatctagtag
     1381 accatttgct gtaatcgacc ttggattgaa gagagagatg attggtgatt tatccactga
     1441 aatgattcca cactttttgg aaagtttcgc ggaggcggcc agaattactt tgcatgttga
     1501 ttgtctgaga ggtttcaacg atcaccacag aagtgagagt gcgttcaagg ctttggctgt
     1561 tgccataaga gaagctattt ctagcaatgg caccaatgac gttccctcaa ccaaaggtgt
     1621 tttgatgtga tcagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta
     1681 tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt
     1741 tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt aatatcatgc
     1801 gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta acgccgccat
     1861 ccagtttaaa cgagctcgaa ttcatcgatg atatcagatc cactagtggc ctatgcggcc
     1921 gcggatctgc cggtctccct atagtgagtc gtattaattt cgataagcca ggttaacctg
     1981 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     2041 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2101 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2161 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2221 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2281 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2341 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2401 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2461 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2521 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2581 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2641 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2701 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2761 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2821 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2881 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2941 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3001 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3061 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3121 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3181 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3241 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3301 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3361 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3421 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3481 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3541 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3601 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3661 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3721 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3781 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3841 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3901 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3961 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     4021 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     4081 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     4141 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     4201 gtactgagag tgcaccatat ggacatattg tcgttagaac gcggctacaa ttaatacata
     4261 accttatgta tcatacacat acgatttagg tgacactata ga
//
