LOCUS Exported 4546 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Truncated C-terminal AID degron cassette with HygR marker for S. cerevisiae. ACCESSION . VERSION . KEYWORDS pHyg-AID(1-114) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4546) 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 4546) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4546 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..19 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1..18 /label=SP6 /note="SP6 promoter, forward primer" promoter 511..854 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 855..1883 /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(1907..1925) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" terminator 1937..2126 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(2200..2219) /label=T7 /note="T7 promoter, forward primer" promoter complement(2201..2219) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2306..2323) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2477..3065) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2557..2576) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3236..4096) /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 3859..3878 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4097..4201) /gene="bla" /label=AmpR promoter primer_bind 4269..4287 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(4325..4347) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 4447..4466 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 atttaggtga cactatagaa cgcggccgcc agctgaagct tcgtacgctg caggtcgacg 61 gtgcaggcgc cggggctggt gctggggcag gtaccatgat gggcagtgtc gagctgaatc 121 tgagggagac tgagctgtgt cttggtcttc ccggtggaga tacagtggct ccggtaaccg 181 gaaacaagag agggttctca gagacggttg atctgaagct aaatctgaat aatgagcctg 241 caaacaagga aggatctacg actcatgacg tcgtgacttt tgattccaag gagaagagtg 301 cttgtcctaa agatccagcc aaacctccgg ccaaggcaca agttgtggga tggccaccgc 361 ttgatttttg gcaggaaacc atcacgttct tccggtatga tctcaccggt ggcccggagg 421 cggcggcgtt cgtgaagtga tcacccgggt taattaaggc gcgccagatc tgtttagctt 481 gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac 541 agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata 601 catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa 661 aattacggct cctcgctgcg gacctgcgag cagggaaacg ctcccctcac agacgcgttg 721 aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg 781 ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc 841 gaacataaac aaccatgggt aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc 901 tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc 961 gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg 1021 atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc 1081 cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg 1141 cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg 1201 tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc 1261 cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg 1321 ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg 1381 cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg 1441 tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca 1501 ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct 1561 ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg 1621 agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct 1681 atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg 1741 caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg 1801 ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca 1861 ctcgtccgag ggcaaaggaa taatctcgag tcatgtaatt agttatgtca cgcttacatt 1921 cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct 1981 aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa 2041 tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc 2101 ttgagaaggt tttgggacgc tcgaaggctt taacgagctc gaattcatcg atgatatcag 2161 atccactagt ggcctatgcg gccgcggatc tgccggtctc cctatagtga gtcgtattaa 2221 tttcgataag ccaggttaac ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 2281 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 2341 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 2401 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 2461 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 2521 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 2581 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 2641 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 2701 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 2761 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 2821 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 2881 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 2941 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 3001 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 3061 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 3121 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 3181 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 3241 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 3301 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 3361 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 3421 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 3481 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 3541 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 3601 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 3661 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 3721 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 3781 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 3841 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 3901 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 3961 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 4021 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 4081 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 4141 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 4201 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa 4261 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg 4321 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg 4381 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta 4441 actatgcggc atcagagcag attgtactga gagtgcacca tatggacata ttgtcgttag 4501 aacgcggcta caattaatac ataaccttat gtatcataca catacg //