LOCUS       sequence-191047-        4544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminal AID*-6FLAG degron cassette with HygR marker for S. 
            cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4544)
  AUTHORS   Morawska M, Ulrich HD
  TITLE     An expanded tool kit for the auxin-inducible degron system in 
            budding yeast.
  JOURNAL   Yeast. 2013 Sep;30(9):341-51. doi: 10.1002/yea.2967. Epub 2013 Jul 
            23.
  PUBMED    23836714
REFERENCE   2  (bases 1 to 4544)
  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"; doi: "10.1002/yea.2967"; 
            journalName: "Yeast"; date: "2013-09"; volume: "30"; issue: "9"; 
            pages: "341-51"
COMMENT     Sequence Label: pHyg-AID*-6FLAG
FEATURES             Location/Qualifiers
     source          1..4544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..210
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Morawska and Ulrich, 
                     2013)"
                     /label=AID*
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="PKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEAAAFVK"
     CDS             256..279
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             286..309
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             316..339
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             346..369
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             376..399
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             406..429
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        492..835
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             836..1864
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             836..1864
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      1872..2119
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1888..1906)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2181..2200)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2182..2200)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2287..2304)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2458..3046)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2538..2557)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3217..4008,4009..4077))
                     /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     3840..3859
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4078..4182)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4250..4268
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4306..4328)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4428..4447
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4528..4544,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4528..4544,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggtgcagg cgccggggct
       61 ggtgctgggg caggtacccc taaagatcca gccaaacctc cggccaaggc acaagttgtg
      121 ggatggccac cggtgagatc ataccggaag aacgtgatgg tttcctgcca aaaatcaagc
      181 ggtggcccgg aggcggcggc gttcgtgaag gtatcacccg ggatacatta tacgaagtta
      241 tgcatgctca ctagtgacta caaggacgac gatgacaagt ctagtgacta caaggacgac
      301 gatgacaagt ctagtgacta caaggacgac gatgacaagt ctagtgacta caaggacgac
      361 gatgacaagt ctagtgacta caaggacgac gatgacaagt ctagtgacta caaggacgac
      421 gatgacaagt ctagatgagg cgcgccagat ctgtttagct tgcctcgtcc ccgccgggtc
      481 acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct
      541 caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat
      601 ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc
      661 ggacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc
      721 ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc
      781 cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa caaccatggg
      841 taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
      901 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
      961 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     1021 ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg
     1081 ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     1141 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc
     1201 gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     1261 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     1321 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     1381 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     1441 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     1501 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     1561 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg
     1621 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     1681 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     1741 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg
     1801 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     1861 ataatctcga gtcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc
     1921 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     1981 tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta
     2041 cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg
     2101 ctcgaaggct ttaacgagct cgaattcatc gatgatatca gatccactag tggcctatgc
     2161 ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa gccaggttaa
     2221 cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2281 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2341 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2401 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2461 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2521 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2581 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2641 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2701 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2761 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2821 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2881 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2941 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3001 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3061 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3121 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3181 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3241 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3301 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3361 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3421 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3481 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3541 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3601 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3661 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3721 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3781 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3841 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3901 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3961 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4021 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4081 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4141 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4201 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     4261 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     4321 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     4381 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     4441 gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct acaattaata
     4501 cataacctta tgtatcatac acatacgatt taggtgacac tata
//
