pFA6a-GFP(S65T)-His3MX6 was a gift from John Pringle (Addgene plasmid # 41598 ; http://n2t.net/addgene:41598 ; RRID:Addgene_41598)

Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae. Longtine MS, McKenzie A 3rd, Demarini DJ, Shah NG, Wach A, Brachat A, Philippsen P, Pringle JR. Yeast. 1998 Jul;14(10):953-61. 10.1002/(SICI)1097-0061(199807)14:10 953::AID-YEA293 3.0.CO;2-U PubMed 9717241
LOCUS pFA6a-GFP(S65T)-His3MX6 5546 bp ds-DNA circular SYN 06-AUG-2018DEFINITION synthetic circular DNAACCESSION .VERSION .KEYWORDS pFA6a-GFP(S65T)-His3MX6SOURCE synthetic DNA construct ORGANISM synthetic DNA constructREFERENCE 1 (bases 1 to 5546) AUTHORS Longtine MS, McKenzie A 3rd, Demarini DJ, Shah NG, Wach A, Brachat A, Philippsen P, Pringle JR TITLE Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae. JOURNAL Yeast. 1998 Jul;14(10):953-61. PUBMED 9717241REFERENCE 2 (bases 1 to 5546) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.comFEATURES Location/Qualifiers source 1..5546 /organism= synthetic DNA construct /mol_type= other DNA CDS 61..774 /codon_start=1 /product= Aequoria victoria green fluorescent protein /label=GFP /translation= SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF ICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF VTAAGITHGMDELYK primer_bind complement(88..116) /label=GFP-R /note= GFP, reverse primer. Does NOT anneal to EGFP primer_bind 711..731 /label=GFP-F /note= GFP, forward primer. Does NOT anneal to EGFP terminator 795..982 /gene= S. cerevisiae ADH1 /label=ADH1 terminator /note= transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene primer_bind complement(850..874) /label=pHybLex-R /note= ADH terminator, reverse primer promoter 1057..1400 /label=TEF promoter /note= Ashbya gossypii TEF promoter terminator 2103..2300 /label=TEF terminator /note= Ashbya gossypii TEF terminator primer_bind complement(2404..2423) /label=T7 /note= T7 promoter, forward primer promoter complement(2405..2423) /label=T7 promoter /note= promoter for bacteriophage T7 RNA polymerase primer_bind complement(2510..2527) /label=L4440 /note= L4440 vector, forward primer rep_origin complement(2681..3269) /direction=LEFT /label=ori /note= high-copy-number ColE1/pMB1/pBR322/pUC origin of replication primer_bind complement(2761..2780) /label=pBR322ori-F /note= pBR322 origin, forward primer CDS complement(3440..4300) /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 4063..4082 /label=Amp-R /note= Ampicillin resistance gene, reverse primer promoter complement(4301..4405) /gene= bla /label=AmpR promoter mobile_element 4466..5233 /mobile_element_type= insertion sequence:IS1 /note= prokaryotic transposable element primer_bind 5250..5268 /label=pBRforEco /note= pBR322 vectors, upsteam of EcoRI site, forward primer primer_bind complement(5306..5328) /label=pGEX 3 /note= pGEX vectors, reverse primer primer_bind 5428..5447 /label=pRS-marker /note= pRS vectors, use to sequence yeast selectable marker promoter 5528..5546 /label=SP6 promoter /note= promoter for bacteriophage SP6 RNA polymerase primer_bind 5528..5545 /label=SP6 /note= SP6 promoter, forward primer ORIGIN 1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac 61 agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat 121 gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa 181 cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc 241 actactttca cttatggtgt tcaatgcttt tcaagatacc cagatcatat gaaacggcat 301 gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa 361 gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat 421 agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg 481 gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc 541 aaagttaact tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat 601 tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg 661 tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt 721 gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa atagggcgcg 781 ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa 841 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat 901 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc 961 ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata tagatctgtt 1021 tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc 1081 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag 1141 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga 1201 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac 1261 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca 1321 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc 1381 acatccgaac ataaacaacc atggcagaac cagcccaaaa aaagcaaaaa caaactgttc 1441 aggagcgcaa ggcgtttatc tcccgtatca ctaatgaaac taaaattcaa atcgctattt 1501 cgctgaatgg tggttatatt caaataaaag attcgattct tcctgcaaag aaggatgacg 1561 atgtagcttc ccaagctact cagtcacagg tcatcgatat tcacacaggt gttggctttt 1621 tggatcatat gatccatgcg ttggcaaaac actctggttg gtctcttatt gttgaatgta 1681 ttggtgacct gcacattgac gatcaccata ctaccgaaga ttgcggtatc gcattagggc 1741 aagcgttcaa agaagcaatg ggtgctgtcc gtggtgtaaa aagattcggt actgggttcg 1801 caccattgga tgaggcgcta tcacgtgccg tagtcgattt atctagtaga ccatttgctg 1861 taatcgacct tggattgaag agagagatga ttggtgattt atccactgaa atgattccac 1921 actttttgga aagtttcgcg gaggcggcca gaattacttt gcatgttgat tgtctgagag 1981 gtttcaacga tcaccacaga agtgagagtg cgttcaaggc tttggctgtt gccataagag 2041 aagctatttc tagcaatggc accaatgacg ttccctcaac caaaggtgtt ttgatgtgaa 2101 gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata 2161 ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca 2221 tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt 2281 gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca gtttaaacga 2341 gctcgaattc atcgatgata tcagatccac tagtggccta tgcggccgcg gatctgccgg 2401 tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat taatgaatcg 2461 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 2521 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 2581 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 2641 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 2701 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 2761 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 2821 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 2881 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 2941 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 3001 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 3061 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 3121 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 3181 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 3241 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 3301 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 3361 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 3421 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 3481 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 3541 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 3601 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 3661 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 3721 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 3781 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 3841 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 3901 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 3961 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 4021 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 4081 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 4141 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 4201 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 4261 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 4321 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 4381 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 4441 aaaccattat tatcatgaca ttaacggtaa tgactccaac ttattgatag tgttttatgt 4501 tcagataatg cccgatgact ttgtcatgca gctccaccga ttttgagaac gacagcgact 4561 tccgtcccag ccgtgccagg tgctgcctca gattcaggtt atgccgctca attcgctgcg 4621 tatatcgctt gctgattacg tgcagctttc ccttcaggcg ggattcatac agcggccagc 4681 catccgtcat ccatatcacc acgtcaaagg gtgacagcag gctcataaga cgccccagcg 4741 tcgccatagt gcgttcaccg aatacgtgcg caacaaccgt cttccggaga ctgtcatacg 4801 cgtaaaacag ccagcgctgg cgcgatttag ccccgacata gccccactgt tcgtccattt 4861 ccgcgcagac gatgacgtca ctgcccggct gtatgcgcga ggttaccgac tgcggcctga 4921 gttttttaag tgacgtaaaa tcgtgttgag gccaacgccc ataatgcggg ctgttgcccg 4981 gcatccaacg ccattcatgg ccatatcaat gattttctgg tgcgtaccgg gttgagaagc 5041 ggtgtaagtg aactgcagtt gccatgtttt acggcagtga gagcagagat agcgctgatg 5101 tccggcggtg cttttgccgt tacgcaccac cccgtcagta gctgaacagg agggacagct 5161 gatagaaaca gaagccactg gagcacctca aaaacaccat catacactaa atcagtaagt 5221 tggcagcatc accgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg 5281 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag 5341 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg 5401 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc 5461 atatggacat attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac 5521 acatacgatt taggtgacac tataga