LOCUS       sequence-495243-        4101 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 4101)
  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 4101)
  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-klTRP1
FEATURES             Location/Qualifiers
     source          1..4101
                     /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"
     primer_bind     complement(1736..1755)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1737..1755)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1842..1859)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2013..2601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2093..2112)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2772..3563,3564..3632))
                     /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     3395..3414
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3633..3737)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3805..3823
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3861..3883)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3983..4002
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4083..4101
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4083..4100
                     /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 ggccagcaaa gtggaacgat
      601 cattcactat atatatatca atttatatat acgtatgtgt aattgaagaa agatacgttt
      661 tttcctctat tgagaggcct gctggatgaa tagctttacc ttttctaaat ccttgatacc
      721 atcagtctct actcctccac ttacatcgac accgatcgca tttggtaaca tattaatggc
      781 aacagaaacg ttatcaggat tcaatccacc agcgataatg aattttatct cgggatgact
      841 tgcagaccaa ctggaaattg cactccaatt caatttctca ccagtgccac cttcaccaga
      901 atcgaacaac gtcagcacat tgtctacgtg ttcatacagg tccagtagta attcacaatc
      961 ctgtgggaac tggaacctct taatgattgg aattgaagat gggatcaaag atctgtattc
     1021 tttaatatct tcatctccat gtaattgtat cacatctaga ttatattcgt ggtacagttg
     1081 aaggacatca tcaacggact gatttctaaa caccccgact agtttagtac ctttcacgtt
     1141 ctcttgttgg tgaactgcag ttgaaatacc tttcgcaaca gatgagtcaa tggttctttt
     1201 cctaccggga acacaaatga tacctaagta atcagcacca tcatccacag cagtctttgc
     1261 agcttcaacg gtttgcaaac cacacacttt aacgagcatc acagtagatt tcgcaagagg
     1321 gtagctcgct cagagtaccc aagtaaatga ttagtaaact gatgtttgat agttcaattt
     1381 ttcaatgaaa taaccttata ttaaaattga tattactatt atacaaaaat aaagaataaa
     1441 ggatttgagt ttatacataa aataccatta ttatttgttc agtgagagat accggggtat
     1501 atgggatgtg tgtagtgata ccatgcaatc atgtatcaaa catggaagaa gtgaaggggg
     1561 agaaagaaac ttgaaaaaaa gaaaaccatt tgaaaagagg aaaagtatct aattaagatc
     1621 tttcacctct tactgtcgat tcgatactaa cgccgccatc cagtttaaac gagctcgaat
     1681 tcatcgatga tatcagatcc actagtggcc tatgcggccg cggatctgcc ggtctcccta
     1741 tagtgagtcg tattaatttc gataagccag gttaacctgc attaatgaat cggccaacgc
     1801 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     1861 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1921 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1981 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2041 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2101 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2161 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2221 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2281 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2341 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2401 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2461 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2521 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2581 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2641 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2701 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2761 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2821 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2881 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2941 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3001 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3061 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3121 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3181 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3241 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3301 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3361 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3421 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     3481 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     3541 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3601 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     3661 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3721 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     3781 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     3841 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     3901 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     3961 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     4021 gacatattgt cgttagaacg cggctacaat taatacataa ccttatgtat catacacata
     4081 cgatttaggt gacactatag a
//
