LOCUS Exported 8110 bp ds-DNA circular SYN 01-APR-2022 DEFINITION C-SWAT compatible vector for C-terminal tagging with SMASh Degron. (Protease Cleavage Site: DEMEECSKHL).. ACCESSION . VERSION . KEYWORDS C-SWAT SMASh SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8110) TITLE A Suite of New C-SWAT Strain Construction Vectors Enables Comparative Evaluation of Essential Gene Perturbation Tools in Yeast JOURNAL Unpublished REFERENCE 2 (bases 1 to 8110) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 1, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8110 /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" CDS 440..1774 /codon_start=1 /gene="S. cerevisiae MET17" /gene_synonym="MET15; MET25" /product="O-acetylserine and O-acetylhomoserine sulfhydrylase, required for sulfur amino acid synthesis" /label=MET17 /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG FGGVLSFGVKDLPNADKETDPFELSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP" rep_origin complement(1879..2334) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2016..2037) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2228..2247 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2460..2482 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2474..2491 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2475..2491 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2501..2520 /label=T7 /note="T7 promoter, forward primer" promoter 2501..2519 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 2544..2560 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 2545..2561 /label=KS primer /note="common sequencing primer, one of multiple similar variants" CDS 2715..2738 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" gene 3775..5131 /label=kanMX /note="yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)" promoter 3775..4118 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 4119..4928 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(4186..4205) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" terminator 4934..5131 /label=TEF terminator /note="Ashbya gossypii TEF terminator" primer_bind complement(5337..5357) /label=T3 /note="T3 promoter, forward primer" promoter complement(5337..5355) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(5376..5392) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5376..5392) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5389..5411) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5400..5416 /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(5424..5454) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5469..5490 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5607..5624) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5778..6366) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5858..5877) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6537..7397) /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 7160..7179 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7398..7502) /gene="bla" /label=AmpR promoter misc_feature 7539..8042 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" primer_bind 8083..8101 /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 accacgcttt tcaattcaat tcatcaccat cctcatgaaa actgtgtaac ataataaccg 241 aagtgtcgaa aaggtggcac cttgtccaat tgaacacgct cgatgaaaaa aataagatat 301 atataaggtt aagtaaagcg tctgttagaa aggaagtttt tcctttttct tgctctcttg 361 tcttttcatc tactatttcc ttcgtgtaat acagggtcgt cagatacata gatacaattc 421 tattaccccc atccatacaa tgccatctca tttcgatact gttcaactac acgccggcca 481 agagaaccct ggtgacaatg ctcacagatc cagagctgta ccaatttacg ccaccacttc 541 ttatgttttc gaaaactcta agcatggttc gcaattgttt ggtctagaag ttccaggtta 601 cgtctattcc cgtttccaaa acccaaccag taatgttttg gaagaaagaa ttgctgcctt 661 agaaggtggt gctgctgctt tggctgtttc ctccggtcaa gccgctcaaa cccttgccat 721 ccaaggtttg gcacacactg gtgacaacat cgtttccact tcttacttat acggtggtac 781 ttataaccag ttcaaaatct cgttcaaaag atttggtatc gaggctagat ttgttgaagg 841 tgacaatcca gaagaattcg aaaaggtctt tgatgaaaga accaaggctg tttatttgga 901 aaccattggt aatccaaagt acaatgttcc ggattttgaa aaaattgttg caattgctca 961 caaacacggt attccagttg tcgttgacaa cacatttggt gccggtggtt acttctgtca 1021 gccaattaaa tacggtgctg atattgtaac acattctgct accaaatgga ttggtggtca 1081 tggtactact atcggtggta ttattgttga ctctggtaag ttcccatgga aggactaccc 1141 agaaaagttc cctcaattct ctcaacctgc cgaaggatat cacggtacta tctacaatga 1201 agcctacggt aacttggcat acatcgttca tgttagaact gaactattaa gagatttggg 1261 tccattgatg aacccatttg cctctttctt gctactacaa ggtgttgaaa cattatcttt 1321 gagagctgaa agacacggtg aaaatgcatt gaagttagcc aaatggttag aacaatcccc 1381 atacgtatct tgggtttcat accctggttt agcatctcat tctcatcatg aaaatgctaa 1441 gaagtatcta tctaacggtt tcggtggtgt cttatctttc ggtgtaaaag acttaccaaa 1501 tgccgacaag gaaactgacc cattcgaact ttctggtgct caagttgttg acaatttaaa 1561 gcttgcctct aacttggcca atgttggtga tgccaagacc ttagtcattg ctccatactt 1621 cactacccac aaacaattaa atgacaaaga aaagttggca tctggtgtta ccaaggactt 1681 aattcgtgtc tctgttggta tcgaatttat tgatgacatt attgcagact tccagcaatc 1741 ttttgaaact gttttcgctg gccaaaaacc atgagtgtgc gtgatgagtt gtaaaattat 1801 gtataaacct actttctctc acaagtactg cggtgtgaaa taccgcacag atgcgtaagg 1861 agaaaatacc gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt 1921 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat 1981 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat 2041 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac 2101 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc 2161 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga 2221 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca 2281 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc 2341 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg 2401 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc 2461 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg 2521 cgaattgggt accgggcccc ccctcgaggt cgacggtatc gataagcttg atatcgaatt 2581 cctgcagcta tattaccctg ttatccctac gtacgctgca ggtcgacggt ggcggttctg 2641 gcggtggcgg atccgacgaa atggaagaat gtagtaagca cttacctggt gcaggctcat 2701 ctggcgatat catggactac aaggacgacg atgataaggg ttcttctgga acaggttcag 2761 gttcaggtac aagtgcaccc attactgctt atgctcagca aacaagaggc ctattgggtt 2821 gtattattac tagtttgacg ggcagagaca aaaaccaggt agaaggtgaa gtgcagatcg 2881 tatccacggc tactcagaca ttcttggcta cttgtattaa tggtgtgtgt tgggcagttt 2941 accacggtgc gggaactaga actattgcct ctcctaaggg tccagtgatc caaatgtata 3001 ctaacgttga tcaagatttg gtcggctggc cagctccaca agggtcaaga agcctaactc 3061 cttgcacctg cggttcttca gatctgtact tagtgacacg tcatgcagat gttataccag 3121 ttagacgtcg tggggacagt agaggttcac tgctgtcacc acgtccaatt agttacctta 3181 aaggtagttc tggcggtccc ctgttgtgtc ctgcaggtca tgcagtagga ttgtttagag 3241 ccgccgtctg caccagaggg gttgcaaaag ctgtggattt tattcctgtt gaaaatcttg 3301 aaacgactat gagatcacct gtctttaccg ataatagttc cccaccagct gtgactctta 3361 cacatccgat caccaaaatc gatactaaat acattatgac ctgtatgtct gccgatttag 3421 aagtagtaac atctacatgg gttttagtcg gtggggtttt agctgcccta gccgcatatt 3481 gcttgtccac aggttgcgtt gttatagttg gcaggatcgt tttgagcgga aaataaagtt 3541 cttctttgag atatcgattg aacgagctcg aattcatcga tctatattac cctgttatcc 3601 ctagcgtaac tatgccaaat gacaataaga cacccaatag gtccagcact cccaagttta 3661 ctaaaaaacc agtaaccccg aatgataaaa ttcctgaaag acggatcccc gggttaatta 3721 aggcgcgcca gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg 3781 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac 3841 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt 3901 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga 3961 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata 4021 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta 4081 ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaaggaa aagactcacg 4141 tttcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc 4201 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc 4261 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg 4321 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta 4381 ctcctgatga tgcatggtta ctcaccactg cgatccccgg caaaacagca ttccaggtat 4441 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc 4501 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg 4561 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc 4621 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac 4681 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga 4741 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg 4801 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa 4861 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt 4921 ttttctaatc agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata 4981 gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt 5041 tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt 5101 caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc 5161 agtttaaacg agctcgaatt ctgatggaaa aagattttca agtgaataag gagataaaac 5221 cgtattgact ttttatcatt ctacaaaaga ccagtaacga agtgtaccct gctatattac 5281 cctgttatcc ctagcgtaac tcggccgcca ccgcggtgga gctccagctt ttgttccctt 5341 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 5401 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 5461 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 5521 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 5581 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 5641 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 5701 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 5761 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 5821 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 5881 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 5941 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 6001 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 6061 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 6121 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 6181 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 6241 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 6301 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 6361 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 6421 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 6481 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 6541 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 6601 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 6661 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 6721 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 6781 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 6841 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 6901 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 6961 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 7021 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 7081 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 7141 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 7201 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 7261 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 7321 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 7381 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 7441 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 7501 cgcacatttc cccgaaaagt gccacctggg tccttttcat cacgtgctat aaaaataatt 7561 ataatttaaa ttttttaata taaatatata aattaaaaat agaaagtaaa aaaagaaatt 7621 aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac tctaggggga tcgccaacaa 7681 atactacctt ttatcttgct cttcctgctc tcaggtatta atgccgaatt gtttcatctt 7741 gtctgtgtag aagaccacac acgaaaatcc tgtgatttta cattttactt atcgttaatc 7801 gaatgtatat ctatttaatc tgcttttctt gtctaataaa tatatatgta aagtacgctt 7861 tttgttgaaa ttttttaaac ctttgtttat ttttttttct tcattccgta actcttctac 7921 cttctttatt tactttctaa aatccaaata caaaacataa aaataaataa acacagagta 7981 aattcccaaa ttattccatc attaaaagat acgaggcgcg tgtaagttac aggcaagcga 8041 tccgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg 8101 ccctttcgtc //