LOCUS       sequence-495240-        5318 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 5318)
  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 5318)
  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-hphNT1
FEATURES             Location/Qualifiers
     source          1..5318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1004..1191
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1059..1083)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1266..1609
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1610..2638
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSNLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2646..2893
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2662..2680)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2955..2974)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2956..2974)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3061..3078)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3232..3820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3312..3331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3991..4782,4783..4851))
                     /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     4614..4633
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4852..4956)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5024..5042
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5080..5102)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5202..5221
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5302..5318,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5302..5318,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acggcttctc tgagaccgtg gacctgatgc tgaacctgca gtccaataag gagggctctg
      121 tggatctgaa gaacgtgagc gccgtgccta aggagaagac cacactgaag gacccatcca
      181 agccccctgc caaggcacag gtggtgggat ggccacccgt gcggaactac agaaagaata
      241 tgatgaccca gcagaagaca agctccggtt ctggtgtttc taagggtgaa gaagacaaca
      301 tggcttcttt gccagctact cacgaattgc acatcttcgg ttctatcaac ggtgttgact
      361 tcgacatggt tggtcaaggt actggtaacc caaacgacgg ttacgaagaa ttgaacttga
      421 agtctactaa gggtgacttg caattctctc catggatctt ggttccacac atcggttacg
      481 gtttccacca atacttgcca tacccagacg gtatgtctcc attccaagct gctatggttg
      541 acggttctgg ttaccaagtt cacagaacta tgcaattcga agacggtgct tctttgactg
      601 ttaactacag atacacttac gaaggttctc acatcaaggg tgaagctcaa gttaagggta
      661 ctggtttccc agctgacggt ccagttatga ctaactcttt gactgctgct gactggtgta
      721 gatctaagaa gacttaccca aacgacaaga ctatcatctc tactttcaag tggtcttaca
      781 ctactggtaa cggtaagaga tacagatcta ctgctagaac tacttacact ttcgctaagc
      841 caatggctgc taactacttg aagaaccaac caatgtacgt tttcagaaag actgaattga
      901 agcactctaa gactgaattg aacttcaagg aatggcaaaa ggctttcact gacgttatgg
      961 gtatggacga attgtacaag taaggcgcgc cacttctaaa taagcgaatt tcttatgatt
     1021 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     1081 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     1141 gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg gcagatccgc
     1201 tagggataac agggtaatat agatctgttt agcttgcctc gtccccgccg ggtcacccgg
     1261 ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg
     1321 catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat
     1381 ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcggacct
     1441 gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt
     1501 agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta
     1561 aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca tgggtaaaaa
     1621 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     1681 caacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     1741 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     1801 ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt
     1861 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     1921 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc
     1981 tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     2041 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     2101 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     2161 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     2221 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     2281 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     2341 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg
     2401 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     2461 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     2521 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
     2581 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaataatc
     2641 tcgagtcatg taattagtta tgtcacgctt acattcacgc cctcccccca catccgctct
     2701 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt tttttatagt
     2761 tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacagacg
     2821 cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg gacgctcgaa
     2881 ggctttaacg agctcgaatt catcgatgat atcagatcca ctagtggcct atgcggccgc
     2941 ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca
     3001 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3061 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3121 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3181 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3241 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3301 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3361 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3421 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3481 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3541 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3601 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3661 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3721 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3781 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3841 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3901 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3961 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagcg aggcacctat
     4021 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4081 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4141 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4201 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4261 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4321 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4381 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4441 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4501 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4561 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4621 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4681 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4741 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4801 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4861 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4921 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4981 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     5041 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     5101 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     5161 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     5221 actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac
     5281 cttatgtatc atacacatac gatttaggtg acactata
//
