LOCUS       sequence-494870-        4129 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 4129)
  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 4129)
  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-klTRP1
FEATURES             Location/Qualifiers
     source          1..4129
                     /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"
     primer_bind     complement(1764..1783)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1765..1783)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1870..1887)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2041..2629)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2121..2140)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2800..3591,3592..3660))
                     /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     3423..3442
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3661..3765)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3833..3851
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3889..3911)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4011..4030
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4111..4129
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4111..4128
                     /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 ccagcaaagt ggaacgatca ttcactatat atatatcaat ttatatatac
      661 gtatgtgtaa ttgaagaaag atacgttttt tcctctattg agaggcctgc tggatgaata
      721 gctttacctt ttctaaatcc ttgataccat cagtctctac tcctccactt acatcgacac
      781 cgatcgcatt tggtaacata ttaatggcaa cagaaacgtt atcaggattc aatccaccag
      841 cgataatgaa ttttatctcg ggatgacttg cagaccaact ggaaattgca ctccaattca
      901 atttctcacc agtgccacct tcaccagaat cgaacaacgt cagcacattg tctacgtgtt
      961 catacaggtc cagtagtaat tcacaatcct gtgggaactg gaacctctta atgattggaa
     1021 ttgaagatgg gatcaaagat ctgtattctt taatatcttc atctccatgt aattgtatca
     1081 catctagatt atattcgtgg tacagttgaa ggacatcatc aacggactga tttctaaaca
     1141 ccccgactag tttagtacct ttcacgttct cttgttggtg aactgcagtt gaaatacctt
     1201 tcgcaacaga tgagtcaatg gttcttttcc taccgggaac acaaatgata cctaagtaat
     1261 cagcaccatc atccacagca gtctttgcag cttcaacggt ttgcaaacca cacactttaa
     1321 cgagcatcac agtagatttc gcaagagggt agctcgctca gagtacccaa gtaaatgatt
     1381 agtaaactga tgtttgatag ttcaattttt caatgaaata accttatatt aaaattgata
     1441 ttactattat acaaaaataa agaataaagg atttgagttt atacataaaa taccattatt
     1501 atttgttcag tgagagatac cggggtatat gggatgtgtg tagtgatacc atgcaatcat
     1561 gtatcaaaca tggaagaagt gaagggggag aaagaaactt gaaaaaaaga aaaccatttg
     1621 aaaagaggaa aagtatctaa ttaagatctt tcacctctta ctgtcgattc gatactaacg
     1681 ccgccatcca gtttaaacga gctcgaattc atcgatgata tcagatccac tagtggccta
     1741 tgcggccgcg gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt
     1801 taacctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1861 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1921 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1981 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2041 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2101 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2161 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2221 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2281 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2341 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2401 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2461 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2521 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2581 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2641 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2701 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2761 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2821 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2881 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2941 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3001 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3061 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3121 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3181 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3241 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3301 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3361 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3421 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3481 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3541 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3601 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3661 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3721 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3781 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     3841 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     3901 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     3961 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     4021 gcagattgta ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta
     4081 atacataacc ttatgtatca tacacatacg atttaggtga cactataga
//
