LOCUS Exported 5432 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pFA6a-kanMX6-PGAL1-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5432) 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 9717241 REFERENCE 2 (bases 1 to 5432) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5432 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(1..19) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2..19) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(100..119) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 219..241 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(279..297) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 365..469 /gene="bla" /label=AmpR promoter CDS 470..1330 /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 complement(688..707) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1501..2089 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1990..2009 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2243..2260 /label=L4440 /note="L4440 vector, forward primer" primer_bind 2347..2366 /label=T7 /note="T7 promoter, forward primer" promoter 2347..2365 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" gene complement(2470..3826) /label=kanMX /note="yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)" terminator 2470..2667 /label=TEF terminator /note="Ashbya gossypii TEF terminator" CDS complement(2673..3482) /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 3396..3415 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" promoter complement(3483..3826) /label=TEF promoter /note="Ashbya gossypii TEF promoter" promoter 3960..4401 /gene="S. cerevisiae GAL1" /label=GAL1 promoter /note="inducible promoter, regulated by Gal4" protein_bind 3960..4077 /label=UAS /bound_moiety="Gal4" /note="upstream activating sequence mediating Gal4-dependent induction" primer_bind 4372..4395 /label=GAL1 /note="S. cerevisiae GAL1 promoter, forward primer" CDS 4436..5149 /codon_start=1 /product="Aequoria victoria green fluorescent protein" /label=GFP /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF ICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF VTAAGITHGMDELYK" primer_bind complement(4463..4491) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 5086..5106 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" terminator 5170..5357 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(5225..5249) /label=pHybLex-R /note="ADH terminator, reverse primer" ORIGIN 1 tctatagtgt cacctaaatc gtatgtgtat gatacataag gttatgtatt aattgtagcc 61 gcgttctaac gacaatatgt ccatatggtg cactctcagt acaatctgct ctgatgccgc 121 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 181 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 241 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt 301 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa 361 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 421 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca 481 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca 541 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta 601 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt 661 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc 721 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc 781 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc 841 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa 901 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga 961 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat 1021 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca 1081 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc 1141 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat 1201 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag 1261 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa 1321 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca 1381 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc 1441 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 1501 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 1561 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 1621 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt 1681 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 1741 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 1801 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 1861 ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg 1921 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 1981 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 2041 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 2101 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 2161 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 2221 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat 2281 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaggttaa cctggcttat 2341 cgaaattaat acgactcact atagggagac cggcagatcc gcggccgcat aggccactag 2401 tggatctgat atcatcgatg aattcgagct cgtttaaact ggatggcggc gttagtatcg 2461 aatcgacagc agtatagcga ccagcattca catacgattg acgcatgata ttactttctg 2521 cgcacttaac ttcgcatctg ggcagatgat gtcgaggcga aaaaaaatat aaatcacgct 2581 aacatttgat taaaatagaa caactacaat ataaaaaaac tatacaaatg acaagttctt 2641 gaaaacaaga atctttttat tgtcagtact gattagaaaa actcatcgag catcaaatga 2701 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt 2761 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct 2821 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg 2881 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta 2941 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc 3001 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg 3061 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc 3121 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttg 3181 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg 3241 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca 3301 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac 3361 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat 3421 aaatcagcat ccatgttgga atttaatcgc ggcctcgaaa cgtgagtctt ttccttaccc 3481 atggttgttt atgttcggat gtgatgtgag aactgtatcc tagcaagatt ttaaaaggaa 3541 gtatatgaaa gaagaacctc agtggcaaat cctaaccttt tatatttctc tacaggggcg 3601 cggcgtgggg acaattcaac gcgtctgtga ggggagcgtt tccctgctcg caggtctgca 3661 gcgaggagcc gtaatttttg cttcgcgccg tgcggccatc aaaatgtatg gatgcaaatg 3721 attatacatg gggatgtatg ggctaaatgt acgggcgaca gtcacatcat gcccctgagc 3781 tgcgcacgtc aagactgtca aggagggtat tctgggcctc catgtcgctg gccgggtgac 3841 ccggcgggga cgaggcaagc taaacagatc tgtaaagagc cccattatct tagcctaaaa 3901 aaaccttctc tttggaactt tcagtaatac gcttaactgc tcattgctat attgaagtac 3961 ggattagaag ccgccgagcg ggtgacagcc ctccgaagga agactctcct ccgtgcgtcc 4021 tcgtcttcac cggtcgcgtt cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac 4081 aataaagatt ctacaatact agcttttatg gttatgaaga ggaaaaattg gcagtaacct 4141 ggccccacaa accttcaaat gaacgaatca aattaacaac cataggatga taatgcgatt 4201 agttttttag ccttatttct ggggtaatta atcagcgaag cgatgatttt tgatctatta 4261 acagatatat aaatgcaaaa actgcataac cactttaact aatactttca acattttcgg 4321 tttgtattac ttcttattca aatgtaataa aagtatcaac aaaaaattgt taatatacct 4381 ctatacttta acgtcaagga gaaaaaaccc ggatctcaaa atgtctttaa ttaacagtaa 4441 aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa 4501 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac 4561 ccttaaattt atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac 4621 tttcacttat ggtgttcaat gcttttcaag atacccagat catatgaaac ggcatgactt 4681 tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga 4741 cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat 4801 cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata 4861 caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt 4921 taacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag accattatca 4981 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac 5041 acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt 5101 tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaatagg gcgcgccact 5161 tctaaataag cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa 5221 ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg 5281 agtaactctt tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt 5341 gaccacacct ctaccggcag atccgctagg gataacaggg taatatagat ccgtcgacct 5401 gcagcgtacg aagcttcagc tggcggccgc gt //