LOCUS       sequence-192008-        6840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal AID*-9myc 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 6840)
  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 6840)
  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)-9myc
FEATURES             Location/Qualifiers
     source          1..6840
                     /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"
     CDS             429..458
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             465..494
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             501..530
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             549..578
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             585..614
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             621..650
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             669..698
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             705..734
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             741..770
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(876..894)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(915..931)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(915..931)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(928..950)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    939..955
                     /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(963..969,970..987,988..993))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1008..1029
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1146..1163)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1317..1905)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1397..1416)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2076..2867,2868..2936))
                     /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     2699..2718
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2937..3041)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3109..3127
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3165..3187)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3287..3306
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3330..3545
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3546..4349
                     /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(4480..4935)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4617..4638)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4829..4848
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5061..5083
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5075..5092
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5076..5092
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5099..5118
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5099..5117
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            5252..6608
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        5252..5595
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             5596..6405
                     /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(5663..5682)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      6411..6608
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     6661..6677
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6661..6677
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
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 tcacccgggg
      421 ctagtggtga acaaaagttg atttctgaag aagatttgaa cggtgaacaa aagctaatct
      481 ccgaggaaga cttgaacggt gaacaaaaat taatctcaga agaagacttg aacggatcct
      541 ctagaggtga acaaaagttg atttctgaag aagatttgaa cggtgaacaa aagctaatct
      601 ccgaggaaga cttgaacggt gaacaaaaat taatctcaga agaagacttg aacggatcct
      661 ctagaggtga acaaaagttg atttctgaag aagatttgaa cggtgaacaa aagctaatct
      721 ccgaggaaga cttgaacggt gaacaaaaat taatctcaga agaagacttg aacggatcca
      781 ctagcctcga ggaagaactt caaattgaat ctaaggactt gggagctggt gcaggcgctg
      841 gagcgggtgc cagagctcgg taccagcttt tgttcccttt agtgagggtt aatttcgagc
      901 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
      961 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     1021 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     1081 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     1141 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1201 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1261 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1321 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1381 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1441 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1501 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1561 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1621 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1681 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1741 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1801 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1861 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1921 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1981 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2041 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     2101 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     2161 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     2221 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     2281 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     2341 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     2401 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     2461 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     2521 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     2581 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     2641 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     2701 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     2761 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     2821 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     2881 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     2941 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     3001 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     3061 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     3121 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     3181 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     3241 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     3301 attgtactga gagtgcacca taccaccttt tcaattcatc attttttttt tattcttttt
     3361 tttgatttcg gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac
     3421 gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat
     3481 gaaattgccc agtattctta acccaactgc acagaacaaa aacctgcagg aaacgaagat
     3541 aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg
     3601 ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc
     3661 gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa
     3721 aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg
     3781 cattatccgc caagtacaat tttttactct tcgaagacag aaaatttgct gacattggta
     3841 atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta
     3901 cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag
     3961 aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc
     4021 tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg
     4081 ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga
     4141 ttatgacacc cggtgtgggt ttagatgaca agggagacgc attgggtcaa cagtatagaa
     4201 ccgtggatga tgtggtctct acaggatctg acattattat tgttggaaga ggactatttg
     4261 caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat
     4321 atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac
     4381 taaactcaca aattagagct tcaatttaat tatatcagtt attaccctat gcggtgtgaa
     4441 ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt
     4501 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4561 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4621 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     4681 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     4741 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     4801 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     4861 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     4921 cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     4981 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     5041 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta
     5101 atacgactca ctatagggcg aattggagct ccaccgcggt ggcggccgcc agctgaagct
     5161 tcgtacgctg caggtcgacg gatccccggg ttaattaagg cgcgccagat ctgtttagct
     5221 tgcctcgtcc ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga
     5281 cagtcttgac gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat
     5341 acatccccat gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa
     5401 aaattacggc tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt
     5461 gaattgtccc cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag
     5521 gttcttcttt catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc
     5581 cgaacataaa caaccatggg taaggaaaag actcacgttt cgaggccgcg attaaattcc
     5641 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     5701 gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     5761 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     5821 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     5881 accactgcga tccccggcaa aacagcattc caggtattag aagaatatcc tgattcaggt
     5941 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     6001 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     6061 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     6121 gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt cactcatggt
     6181 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     6241 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     6301 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     6361 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcagt actgacaata
     6421 aaaagattct tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc
     6481 tattttaatc aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag
     6541 atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg
     6601 ctatactgct gtcgattcga tactaacgcc gccatccagt gtcgaaaacg aggcggccgc
     6661 tctagaacta gtggatccca ttaccgacat ttgggcgcta tacgtgcata tgttcatgta
     6721 tgtatctgta tttaaaacac ttttgtatta tttttcctca tatatgtgta taggtttata
     6781 cggatgattt aattattact tcaccaccct ttatttcagg ctgatatctt agccttgtta
//
