LOCUS Exported 4544 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION C-terminal AID*-6FLAG degron cassette with HygR marker for S. cerevisiae. ACCESSION . VERSION . KEYWORDS pHyg-AID*-6FLAG 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4544 /organism="synthetic DNA construct" /mol_type="other DNA" 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 /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" primer_bind complement(1888..1906) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" terminator 1918..2107 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" 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(3217..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 4528..2 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 4528..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 //