LOCUS       sequence-420146-        7698 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Consitutive expression of myc-E3(1)-CUbo-mCherry-VSV in budding 
            yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7698)
  AUTHORS   Renz C, Asimaki E, Meister C, Albanese V, Petriukov K, Krapoth NC, 
            Wegmann S, Wollscheid HP, Wong RP, Fulzele A, Chen JX, Leon S, 
            Ulrich HD
  TITLE     Ubiquiton-An inducible, linkage-specific polyubiquitylation tool.
  JOURNAL   Mol Cell. 2024 Jan 18;84(2):386-400.e11. doi: 
            10.1016/j.molcel.2023.11.016. Epub 2023 Dec 15.
  PUBMED    38103558
REFERENCE   2  (bases 1 to 7698)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.molcel.2023.11.016"; journalName: "Mol Cell"; date: 
            "2024-01-18- 18"; volume: "84"; issue: "2"; pages: "386-400.e11"
COMMENT     Sequence Label: YIp211-ADH-myc-E3(1)-CUbo-mCherry-VSV
FEATURES             Location/Qualifiers
     source          1..7698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..29)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(183..771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(263..282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(942..1733,1734..1802))
                     /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     1565..1584
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1803..1907)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1975..1993
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2000..2215
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2216..3019
                     /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"
     primer_bind     complement(3137..3159)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3259..3278
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3472..3494
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3486..3503
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3487..3503
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3508..4211
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             4220..4249
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6017..6337
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             6482..7189
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6627..6645)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6895..6914
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             7199..7231
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     terminator      7268..7455
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(7323..7347)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(7479..7495)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7479..7495)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7492..7514)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7503..7519
                     /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(7527..7533,7534..7551,7552..7557))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7572..7593
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
       61 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      601 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      841 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      901 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      961 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1021 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1081 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1141 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1201 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1261 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1321 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1381 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1441 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1501 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1561 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1621 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1681 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1741 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1801 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1861 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1921 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     1981 cgtatcacga ggccagcttt tcaattcatc attttttttt tattcttttt tttgatttcg
     2041 gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga
     2101 gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc
     2161 agtattctta acccaactgc acagaacaaa aacatgcagg aaacgaagat aaatcatgtc
     2221 gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt
     2281 taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa
     2341 ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt
     2401 ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc
     2461 caagtacaat tttttactct tcgaagacag aaaatttgct gacattggta atacagtcaa
     2521 attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca
     2581 cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa
     2641 ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg
     2701 agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt
     2761 tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc
     2821 cggtgtgggt ttagatgaca agggagacgc attgggtcaa cagtatagaa ccgtggatga
     2881 tgtggtctct acaggatctg acattattat tgttggaaga ggactatttg caaagggaag
     2941 ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag
     3001 atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca
     3061 aattagagct tcaatttaat tatatcagtt attacccgcc ctttcgtctc gcgcgtttcg
     3121 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     3181 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     3241 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     3301 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     3361 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     3421 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     3481 gacgttgtaa aacgacggcc agtgaattcc ttttgttgtt tccgggtgta caatatggac
     3541 ttcctctttt ctggcaacca aacccataca tcgggattcc tataatacct tcgttggtct
     3601 ccctaacatg taggtggcgg aggggagata tacaatagaa cagataccag acaagacata
     3661 atgggctaaa caagactaca ccaattacac tgcctcattg atggtggtac ataacgaact
     3721 aatactgtag ccctagactt gatagccatc atcatatcga agtttcacta ccctttttcc
     3781 atttgccatc tattgaagta ataataggcg catgcaactt cttttctttt tttttctttt
     3841 ctctctcccc cgttgttgtc tcaccatatc cgcaatgaca aaaaaatgat ggaagacact
     3901 aaaggaaaaa attaacgaca aagacagcac caacagatgt cgttgttcca gagctgatga
     3961 ggggtatctc gaagcacacg aaactttttc cttccttcat tcacgcacac tactctctaa
     4021 tgagcaacgg tatacggcct tccttccagt tacttgaatt tgaaataaaa aaaagtttgc
     4081 tgtcttgcta tcaagtataa atagacctgc aattattaat cttttgtttc ctcgtcattg
     4141 ttctcgttcc ctttcttcct tgtttctttt tctgcacaat atttcaagct ataccaagca
     4201 tacaatcaac tccggaatgg aacaaaagct tatttctgaa gaagacttgc agattttcgt
     4261 caagactttg accggtaaaa ccataacatt ggaagttgaa tcttccgata ccatcgacaa
     4321 cgttaagtcg cgtattcaag acaaggaagg cattccacct gatcaacaaa gattgatctt
     4381 tgccggtaga cagctcgagg acggtagaac gctgtctgat tacaacattc agcgtgagtc
     4441 caccttacat cttgtgctaa ggctacgtgg cctcatgcag attttcgtca agactttgac
     4501 cggtaaaacc ataacattgg aagttgaatc ttccgatacc atcgacaacg ttaagtcgcg
     4561 tattcaagac aaggaaggca ttccacctga tcaacaaaga ttgatctttg ccggtagaca
     4621 gctcgaggac ggtagaacgc tgtctgatta caacattcag cgtgagtcca ccttacatct
     4681 tgtgctaagg ctacgtggcc tcgcttctgg tgctggcggt tctgaaggtg gcggatctga
     4741 aggtggcaca tctggcgcta caagatctgc ttctggtgct ggtggatctg aaggtggagg
     4801 ttctgaaggt ggtacctctg gtgctacctg tgctgtttgt ggttgggctt tgccacacaa
     4861 cagaatgcaa gctttgacct cttgtgaatg taccatctgt ccagactgtt tcagacaaca
     4921 cttcaccatc gctttgaagg aaaagcacat caccgacatg gtttgtccag cttgtggtag
     4981 accagacttg accgacgaca cccaattgtt gtcttacttc tctaccttgg acatccaatt
     5041 gagagaatct ttggaaccag acgcttacgc tttgttccac aagaagttga ccgaaggtgt
     5101 tttgatgaga gacccaaagt tcttgtggtg tgctcaatgt tctttcggtt tcatctacga
     5161 aagagaacaa ttggaagcta cctgtccaca atgtcaccaa accttctgtg ttagatgtaa
     5221 gagacaatgg gaagaacaac acagaggtag atcatgtgaa gacttccaaa actggaagag
     5281 aatgaacgac ccagaatacc aagctcaagg tttggctatg tacttgcaag aaaacggtat
     5341 cgactgtcca aagtgtaagt tctcttacgc tttggctaga ggtggttgta tgcacttcca
     5401 ctgtacccaa tgtagacacc aattctgttc tggttgttac aacgctttct acgctaagaa
     5461 caagtgtcca gaaccaaact gtagagttaa gaagtctttg cacggtcacc acccaagaga
     5521 ctgtttgttc tacttgagag actggaccgc tttgagattg caaaagttgt tgcaagacaa
     5581 caacgttatg ttcaacaccg aaccaccagc tggtgctaga gctgttccag gtggtggttg
     5641 tagagttatc gaacaaaagg aagttccaaa cggtttgaga gacgaagctt gtggtaagga
     5701 aaccccagct ggttacgctg gtttgtgtca agctcactac aaggaatact tggtttcttt
     5761 gatcaacgct cactctttgg acccagctac cttgtacgaa gttgaagaat tggaaaccgc
     5821 taccgaaaga tacttgcacg ttagaccaca accattggct ggtgaagacc caccagctta
     5881 ccaagctaga ttgttgcaaa agttgaccga agaagttcca ttgggtcaat ctatcccaag
     5941 aagaagaaag ggatccgctt ctggtgctgg tggatctgaa ggtggaggtt ctgaaggtgg
     6001 tacctctggt gctagcggtg ttcaagttga aaccatctct ccaggtgacg gtagaacctt
     6061 cccaaagaga ggtcaaacct gtgttgttca ctacaccggt atgttggaag acggtaagaa
     6121 gttcgactct tctagagaca gaaacaagcc attcaagttc atgttgggta agcaagaagt
     6181 tatcagaggt tgggaagaag gtgttgctca aatgtctgtt ggtcaaagag ctaagttgac
     6241 catctctcca gactacgctt acggtgctac cggtcaccca ggtattatcc caccacacgc
     6301 taccttggtt ttcgacgttg aattgttgaa gttggaaggg tcgaccggta tcccaccaga
     6361 ccaacaaaga ttgatcttcg ctggtaagca attggaagac ggtagaacct tgtctgacta
     6421 caacatccaa aaggaatcta ccttgcactt ggttttgaga ttgagaggtg ttttaattaa
     6481 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     6541 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     6601 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     6661 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     6721 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     6781 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     6841 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     6901 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     6961 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     7021 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     7081 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     7141 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaaat taattaatta
     7201 tactgacata gaaatgaaca gattgggaaa gtagggccct agcggcctgc agccaagcta
     7261 attccgggcg aatttcttat gatttatgat ttttattatt aaataagtta taaaaaaaat
     7321 aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct tattcttgag
     7381 taactctttc ctgtaggtca ggttgctttc tcaggtatag catgaggtcg ctcttattga
     7441 ccacacctct accggcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     7501 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     7561 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     7621 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     7681 ggtttgcgta ttgggcgc
//
