LOCUS Exported 6436 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION S. cerevisiae expression of F-box protein TIR1-9myc under control of the ADH1 promoter, TRP1 marker (for use with the AID degron system). ACCESSION . VERSION . KEYWORDS YIp204-PADH1-atTIR1-9myc SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6436) TITLE Unpublished Ulrich lab AID plasmids REFERENCE 2 (bases 1 to 6436) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6436 /organism="synthetic DNA construct" /mol_type="other DNA" 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(942..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 1995..2096 /gene="S. cerevisiae TRP1" /label=TRP1 promoter CDS 2097..2771 /codon_start=1 /gene="S. cerevisiae TRP1" /product="phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis" /label=TRP1 /note="yeast auxotrophic marker" /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA KK" primer_bind complement(2890..2912) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 3012..3031 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3225..3247 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3239..3256 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3240..3256 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 3554..3950 /gene="S. cerevisiae ADH1" /label=ADH1 promoter /note="promoter for alcohol dehydrogenase 1" CDS 5778..5807 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 5814..5843 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 5850..5879 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 5898..5927 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 5934..5963 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 5970..5999 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 6018..6047 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 6054..6083 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 6090..6119 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" primer_bind complement(6217..6233) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6217..6233) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6230..6252) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6241..6257 /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(6265..6295) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6310..6331 /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 aaggacagta 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 ggccaattcg gtcgaaaaaa gaaaaggaga gggccaagag ggagggcatt 2041 ggtgactatt gagcacgtga gtatacgtga ttaagcacac aaaggcagct tggagtatgt 2101 ctgttattaa tttcacaggt agttctggtc cattggtgaa agtttgcggc ttgcagagca 2161 cagaggccgc agaatgtgca ctagattccg atgctgactt gctgggtatt atatgtgtgc 2221 ccaatagaaa gagaacaatt gacccggtta ttgcaaggaa aatttcaagt cttgtaaaag 2281 catataaaaa tagttcaggc actccgaaat acttggttgg cgtgtttcgt aatcaaccta 2341 aggaggatgt tttggctctg gtcaatgatt acggcattga tatcgtccaa ctgcatggag 2401 atgagtcgtg gcaagaatac caagagttcc tcggtttgcc agttattaaa agactcgtat 2461 ttccaaaaga ctgcaacata ctactcagtg cagcttcaca gaaacctcat tcgtttattc 2521 ccttgtttga ttcagaagca ggtgggacag gtgaactttt ggattggaac tcgatttctg 2581 actgggttgg aaggcaagag agccccgaga gcttacattt tatgttagct ggtggactga 2641 cgccagaaaa tgttggtgat gcgcttagat taaatggcgt tattggtgtt gatgtaagcg 2701 gaggtgtgga gacaaatggt gtaaaagact ctaacaaaat agcaaatttc gtcaaaaatg 2761 ctaagaaata ggttattact gagtagtatt tatttaagta ttgtttgtgc acttgcctgc 2821 aagccttttg aaaagcaagc ataaaagatc gccctttcgt ctcgcgcgtt tcggtgatga 2881 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 2941 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 3001 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat 3061 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg 3121 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 3181 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 3241 taaaacgacg gccagtgaat tcgggtgtac aatatggact tcctcttttc tggcaaccaa 3301 acccatacat cgggattcct ataatacctt cgttggtctc cctaacatgt aggtggcgga 3361 ggggagatat acaatagaac agataccaga caagacataa tgggctaaac aagactacac 3421 caattacact gcctcattga tggtggtaca taacgaacta atactgtagc cctagacttg 3481 atagccatca tcatatcgaa gtttcactac cctttttcca tttgccatct attgaagtaa 3541 taataggcgc atgcaacttc ttttcttttt ttttcttttc tctctccccc gttgttgtct 3601 caccatatcc gcaatgacaa aaaaatgatg gaagacacta aaggaaaaaa ttaacgacaa 3661 agacagcacc aacagatgtc gttgttccag agctgatgag gggtatctcg aagcacacga 3721 aactttttcc ttccttcatt cacgcacact actctctaat gagcaacggt atacggcctt 3781 ccttccagtt acttgaattt gaaataaaaa aaagtttgct gtcttgctat caagtataaa 3841 tagacctgca attattaatc ttttgtttcc tcgtcattgt tctcgttccc tttcttcctt 3901 gtttcttttt ctgcacaata tttcaagcta taccaagcat acaatcaact atctcatata 3961 caactagtat gcagaagcga atagccttgt cgtttccaga agaggtacta gagcatgtgt 4021 tctcgtttat tcagctggat aaggatagga actcagtctc tctggtgtgc aagtcatggt 4081 acgagatcga gcggtggtgc aggaggaaag tcttcatcgg gaactgctac gccgtgagtc 4141 cagcgacggt gattaggagg ttcccgaaag tgagatccgt ggagcttaaa ggaaaacctc 4201 actttgctga ctttaatttg gtacctgacg gatggggagg ttacgtgtat ccatggattg 4261 aggccatgtc ttcgtcttac acgtggcttg aagagataag gctgaagagg atggtggtca 4321 ccgacgattg cttggagctc atagccaagt cttttaagaa ttttaaggtt cttgtgcttt 4381 cttcctgcga aggcttctcc accgatggtc tcgctgctat cgctgccact tgcaggaatc 4441 tgaaagagct tgacttacga gagagtgatg ttgacgacgt tagtggccac tggcttagcc 4501 atttcccaga tacatacact tctttggtat cactcaatat atcttgctta gcatctgagg 4561 tcagtttctc tgctctggaa aggctggtga ctaggtgtcc caatctcaag tctctcaagc 4621 ttaaccgagc tgttccactt gaaaaattgg ctactttact tcaaagagca cctcaattgg 4681 aggaattggg cactggtggg tacactgcag aagtgcgacc agatgtttac tctggtttat 4741 ctgtagcgct ctctgggtgc aaggaattga ggtgcttatc tggattttgg gatgctgttc 4801 ctgcctatct tccagcagtt tattcggttt gcagtcggct tacaactttg aatctgagtt 4861 atgcaacagt ccagagctat gatcttgtca agcttctttg tcaatgccct aaactgcagc 4921 gcctctgggt gcttgactac atcgaggatg ctggtcttga ggtgcttgct tcaacctgca 4981 aggacctacg cgagctgaga gtgtttccgt ccgagccttt tgtcatggaa ccaaatgtgg 5041 cattgacgga acaggggctt gtctccgtct ccatgggctg tccaaaactc gagtcggttc 5101 tctacttctg ccgtcaaatg accaatgctg cattgataac cattgctagg aaccgtccca 5161 acatgactcg cttccgtttg tgcatcattg agccaaaagc cccagactat ctgactctag 5221 agccactgga tattggattt ggagccatag tagagcactg caaggatctc cgtcgcctct 5281 ctctatctgg cctcttgacc gacaaggttt ttgaatacat tgggacatat gccaagaaga 5341 tggaaatgct ctcagtggca tttgcaggag acagtgactt aggcatgcat catgttttgt 5401 ccgggtgcga tagcttgagg aaactagaga taagggactg cccgtttgga gacaaggcgc 5461 ttttggccaa tgcttcaaag ctggagacaa tgcgatccct ttggatgtct tcttgttccg 5521 tgagttttgg agcctgcaag ttactaggac agaagatgcc aaagctgaat gtggaagtca 5581 tcgatgaacg gggtgcaccg gactcgagac cagagagctg ccctgttgag agagtcttca 5641 tataccgaac agtggctggt cctcgatttg acatgcctgg cttcgtctgg aacatggacc 5701 aagactcaac aatgaggttt tccaggcaaa tcattactac taacggatta gtcgactccg 5761 gttctgctgc tagtggtgaa caaaagttga tttctgaaga agatttgaac ggggaacaaa 5821 agctaatctc cgaggaagac ttgaacggtg aacaaaaatt aatctcagaa gaagacttga 5881 acggatcctc tagaggtgaa caaaagttga tttctgaaga agatttgaac ggtgaacaaa 5941 agctaatctc cgaggaagac ttgaacggtg aacaaaaatt aatctcagaa gaagacttga 6001 acggatcctc tagaggtgaa caaaagttga tttctgaaga agatttgaac ggtgaacaaa 6061 agctaatctc cgaggaagac ttgaacggtg aacaaaaatt aatctcagaa gaagacttga 6121 acggatccac tagctaaaaa gtggaacgat cattcaagag atccccgggg atcctctaga 6181 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 6241 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 6301 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 6361 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 6421 tttgcgtatt gggcgc //