LOCUS       sequence-192007-        6298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal AID* degron cassette with CUP1 promoter and KanR marker 
            for S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6298)
  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 6298)
  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: pKan-PCUP1-AID*(N)
FEATURES             Location/Qualifiers
     source          1..6298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..235
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             276..407
                     /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"
     promoter        complement(507..525)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(546..562)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(546..562)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(559..581)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    570..586
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(594..600,601..618,619..624))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    639..660
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(777..794)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(948..1536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1028..1047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1707..2498,2499..2567))
                     /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     2330..2349
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2568..2672)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2740..2758
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2796..2818)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2918..2937
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2961..3176
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3177..3980
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(4111..4566)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4248..4269)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4460..4479
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4692..4714
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4706..4723
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4707..4723
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4730..4749
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4730..4748
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            4883..6239
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        4883..5226
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             5227..6036
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5294..5313)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      6042..6239
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 ctagttagaa aaagacattt ttgctgtcag tcactgtcaa gagattcttt tgctggcatt
       61 tcttctagaa gcaaaaagag cgatgcgtct tttccgctga accgttccag caaaaaagac
      121 taccaacgca atatggattg tcagaatcat ataaaagaga agcaaataac tccttgtctt
      181 gtatcaattg cattataata tcttcttgtt agtgcaatat catatagaag tcatcgaaat
      241 agatattaag aaaaacaaac tgtaacgaat tcatgcctaa agatccagcc aaacctccgg
      301 ccaaggcaca agttgtggga tggccaccgg tgagatcata ccggaagaac gtgatggttt
      361 cctgccaaaa atcaagcggt ggcccggagg cggcggcgtt cgtgaaggta tcaatgctcg
      421 aggaagaact tcaaattgaa tctaaggact tgggagctgg tgcaggcgct ggagcgggtg
      481 ccagagctcg gtaccagctt ttgttccctt tagtgagggt taatttcgag cttggcgtaa
      541 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
      601 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
      661 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
      721 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
      781 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
      841 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
      901 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      961 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1021 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1081 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1141 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1201 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1261 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1321 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1381 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1441 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1501 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1561 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     1621 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1681 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     1741 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1801 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     1861 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1921 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1981 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2041 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2101 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2161 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2221 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2281 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     2341 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2401 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2461 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2521 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     2581 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2641 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     2701 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     2761 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     2821 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     2881 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     2941 agagtgcacc ataccacctt ttcaattcat catttttttt ttattctttt ttttgatttc
     3001 ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
     3061 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
     3121 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
     3181 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
     3241 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
     3301 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
     3361 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
     3421 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
     3481 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
     3541 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
     3601 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
     3661 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
     3721 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     3781 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     3841 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     3901 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     3961 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     4021 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     4081 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat
     4141 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4201 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4261 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4321 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4381 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4441 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4501 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4561 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     4621 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     4681 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc
     4741 actatagggc gaattggagc tccaccgcgg tggcggccgc cagctgaagc ttcgtacgct
     4801 gcaggtcgac ggatccccgg gttaattaag gcgcgccaga tctgtttagc ttgcctcgtc
     4861 cccgccgggt cacccggcca gcgacatgga ggcccagaat accctccttg acagtcttga
     4921 cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac atttagccca tacatcccca
     4981 tgtataatca tttgcatcca tacattttga tggccgcacg gcgcgaagca aaaattacgg
     5041 ctcctcgctg cagacctgcg agcagggaaa cgctcccctc acagacgcgt tgaattgtcc
     5101 ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga ggttcttctt
     5161 tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat ccgaacataa
     5221 acaaccatgg gtaaggaaaa gactcacgtt tcgaggccgc gattaaattc caacatggat
     5281 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     5341 tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     5401 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     5461 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     5521 atccccggca aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     5581 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     5641 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     5701 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     5761 gaaatgcata agcttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     5821 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     5881 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     5941 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     6001 ttgcagtttc atttgatgct cgatgagttt ttctaatcag tactgacaat aaaaagattc
     6061 ttgttttcaa gaacttgtca tttgtatagt ttttttatat tgtagttgtt ctattttaat
     6121 caaatgttag cgtgatttat attttttttc gcctcgacat catctgccca gatgcgaagt
     6181 taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg aatgctggtc gctatactgc
     6241 tgtcgattcg atactaacgc cgccatccag tgtcgaaaac gaggcggccg ctctagaa
//
