LOCUS Exported 7336 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Yeast marker swap plasmid for converting URA3 to ADE2. ACCESSION . VERSION . KEYWORDS M3499 ura3::ADE2 Disruptor Converter SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7336) AUTHORS Voth WP, Jiang YW, Stillman DJ TITLE New 'marker swap' plasmids for converting selectable markers on budding yeast gene disruptions and plasmids. JOURNAL Yeast. 2003 Aug;20(11):985-93. PUBMED 12898713 REFERENCE 2 (bases 1 to 7336) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7336 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 151..170 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 364..386 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 378..395 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 379..395 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 410..625 /gene="S. cerevisiae URA3" /label=URA3 promoter CDS 1232..1243 /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" CDS complement(1542..3257) /codon_start=1 /gene="S. cerevisiae ADE2" /product="phosphoribosylaminoimidazole carboxylase, required for adenine biosynthesis" /label=ADE2 /note="yeast auxotrophic marker" /translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK" promoter complement(3258..3633) /gene="S. cerevisiae ADE2" /label=ADE2 promoter misc_feature 5049..5105 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(5115..5131) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5115..5131) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5128..5150) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5139..5155 /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(5163..5193) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5208..5229 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5346..5363) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5517..6105) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5597..5616) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6276..7136) /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 6899..6918 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7137..7241) /gene="bla" /label=AmpR promoter primer_bind 7309..7327 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttttcaat tcaattcatc 421 attttttttt tattcttttt tttgatttcg gtttccttga aatttttttg attcggtaat 481 ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat 541 atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa 601 aacctgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac 661 tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt 721 gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg 781 tcccaaaatt tgtttactaa aaacacatgt ggatgatccg ggatctcaac gttgttactg 841 gtgtgttgaa gcgatactta agaaagctcc ccaaccctat ctttaccttt caaatatacg 901 agccgttgat gaggttagtt aaatctaaaa aaatgatgga aaacttgcca tttgttggag 961 gaaagttgtc cttagaagca aagaattcgg acacttatat gtcgagcaag agcgctttaa 1021 aaaacatatt ggaagacctt ccaagggaac attatagggt gttaagagta ctgagtgaac 1081 atatagaaaa ggttacacgg tacagtcact ggaatcgaat gacgctttat aatttggctt 1141 tagtttttgc tccaggtttg attcgtgatt tcagtggaga aaaggatatt attgatatga 1201 aagaaagaaa ctatattgtc gcatttatat ttgggaacta caaagatatc ctgacgtagc 1261 gctatcctcg gttctgcatt gagccgcctt atatgaactg tatcgaaacg ttattttttt 1321 aatcgcagac ttaagcaggt aattattcct tgcttcttgt tactggatat gtatgtatgt 1381 ataataagtg atcttatgta tgaaattctt aaaaaaggac acctgtaagc gttgatttct 1441 atgtatgaag tccacatttg atgtaatcat aacaaagcct aaaaaatagg tatatcattt 1501 tataattatt tgctgtacaa gtatatcaat aaacttatat attacttgtt ttctagataa 1561 gcttcgtaac cgacagtttc taacttttgt gctttgacaa gaacttcttc ttcttgcttt 1621 aataaaaact gttccatttt cgttgtataa cttgaatcat aagcgccaag cagtctgaca 1681 gccaacagcg cagcgttcgt actattatta atagcgacgg tagctactgg aacacctcta 1741 ggcatttgca caattgaatg taaagaatct actccatcta gacaagaacc ttttacgggc 1801 acaccgatga caggaagtgg tgtcattgca gccaccatac ctggcaagtg agcagcccca 1861 ccagctccag cgataattgt tttaattcca cgcttgcttg cggaaatagc atatgctgac 1921 atcctatgtg gagttctatg agcagagact attgtcactt caaatggaac gccaaaatct 1981 tttaaaaccg cacatgcggc agacattacc ggcaagtcag agtctgatcc catgatgatt 2041 ccaaccaatg gtttgaccat tgcttccaag tccaactttt gagcgacaga gattttgatt 2101 ggaatatcag ttctacctgt aatgtagttc agcctttgtt cacattccgc catactggag 2161 gcaataatat ttatgtgacc tacttttctg ttaggtctag actcttttcc atataagtac 2221 actgaggaac ctggagtcgc caatgctctt tcgcaagttt ctagctcttt atcttttgta 2281 tgtttgtctc caagaacatt tagcataatg gcgttcgttg taatggtgga gaaagatgtg 2341 aaattctttg gcattggcaa atccaatatt gatctcaaat gagcttcaaa ttgagaagtg 2401 acgcaagcat caatggtata atgtccagag ttgtgaggcc ttggggcaat ttcgttaata 2461 agcaattccc ctgtttctaa atagaacatt tccacaccaa atataccaca accgggaaaa 2521 gatttgattg cattttctgc caacaacttc gccttaagtt gaacggagtc cggaactcta 2581 gcaggcgcat aacataagtc acaaatattg tccttgtgga tagtctctac aattgggtaa 2641 gaaaacacta aaccgttaac agatctcaca atcatgactg ctaattcttt agtaaatggt 2701 gcccattttt cggcgtacaa aggacgatcc ttcagtactt ccaaagcttc cggaatcatt 2761 tccttattct ttacaacgaa gttacctctt ccatcgtatg ccaaagtcct cgacttcaag 2821 acgaatggaa aacccaaatc tcttccaaca ttcaataggg acgtctcact ggcttgttcc 2881 acaggaacac tttgggtaac tgctatacca tttttgatta aatgctcttt ttgaatatat 2941 ttgtcttgta tcaatctgat tgtttctgga gaagggtaaa tttttaattt gggatgtttt 3001 acttgaagat tctttagtgt aggaacatca acatgctcaa tctcaatcgt tagcacatca 3061 catttttcag ctagtttttc gatatcaaga ggattggaaa aggagccatt aacgtggtca 3121 ttggagttgc ttatttgttt ggcaggagaa ttttcagcat ctagtattac cgtcttaatg 3181 ttgagcctgt ttgctgcctc aacaatcata cgtcccaatt gtccccctcc taatatacca 3241 actgttctag aatccatact tgattgtttt gtccgatttt cttgtttttc ttgattgtta 3301 tagtaggatg tacttagaag agagatccaa cgattttacg caccaattta tacatgaaat 3361 gctccataat attgtccatt tagttcttaa taaaaggtca gcaagagtca atcacttagt 3421 attacccggt tcgtagccat gcaacaagag tcatttgtca gcatagctgt aataatcaat 3481 catgacgtaa gaaatgtatc ataattaaaa gttgttaaag atgtcagtgt tatgttggtg 3541 ttacaaaatt ctcggcttct cactaatatt taatatctct taaattttat ctgtctttga 3601 ttcttttaag aaaagttatg tattattcaa gaaaaagtca attccgcatc aaaaggtaaa 3661 atttatataa actgctttaa aatttcatga aactaggcaa cttttcgaaa tgatcttttt 3721 cgagcatgaa gtttctttta taataacctg gtcaaaagct ttcaatatat aatacatttg 3781 gtatttacgg aaatgagatg atatactggt agtgcgctag tcaagttcta attaaattaa 3841 aatgcaaaat agtaatgtca gatggacaca ttgggcgaaa tagctaaccc ataagcatga 3901 atcctgtaac ttatgtaata cgttaatcac gatacggcgt tactagtctt ccttttgagt 3961 ggtttctaat gtagattctt gttgttcgtt aatttgcttc aagccgttta gcattcctga 4021 ttcgtttgaa gatagaggaa cagatgcgcc agcgaaagat gatgatgcaa tggtagagga 4081 agtgttggtc gaatctccct ttttggaaat aaagtgtcgt tttgattcta gataggcttc 4141 tcgcaattga ttgtggagat acttctttaa aagggccata cctatgggat gacattctaa 4201 cctatagagt aatttcgtaa tagcatagca gatttttgac gcccttcttt ctagtagagg 4261 agaagccctt agcttgggtg agatgactga ggatgcgttt gtaaattcgg gataattgga 4321 atttccagta ccctgcccat tacctcccag aatccttata ttagggcgtg tactcagttg 4381 taaaaactgt ctcaatgaag aagattgagc aattctggtc aaatacgttc ttctcctgga 4441 cagagatttt gtgaagggca aatcctgccc actttctttt gtaataaatg tgaggaaaaa 4501 cgccaagata tcaaaactgt aaatttgagt aaaaagatcc cggatccctt ttgatgttag 4561 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca 4621 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa 4681 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag 4741 acgcattggg tcaacagtat agaaccgtgg atgatgtggt ctctacagga tctgacatta 4801 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt 4861 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg 4921 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc 4981 agttattacc cgggaatctc ggtcgtaatg atttttataa tgacgaaaaa aaaaaaattg 5041 gaaagaaaaa gcttgcatgc ctgcaggtcg actctagagg atccccgggt accgagctcg 5101 aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 5161 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 5221 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 5281 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 5341 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 5401 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 5461 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 5521 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 5581 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 5641 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 5701 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 5761 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 5821 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 5881 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 5941 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc 6001 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 6061 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 6121 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 6181 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 6241 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 6301 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 6361 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 6421 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 6481 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 6541 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 6601 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 6661 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 6721 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 6781 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 6841 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 6901 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 6961 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 7021 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 7081 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 7141 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 7201 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 7261 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc 7321 acgaggccct ttcgtc //