LOCUS Exported 4342 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION genomic G11-5FLAG epitope tagging in S. pombe. ACCESSION . VERSION . KEYWORDS pFA6a-G11-5FLAG-kanMX6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4342) AUTHORS Gadaleta MC, Iwasaki O, Noguchi C, Noma K, Noguchi E TITLE New vectors for epitope tagging and gene disruption in Schizosaccharomyces pombe. JOURNAL Biotechniques. 2013 Nov;55(5):257-63. doi: 10.2144/000114100. PUBMED 24215641 REFERENCE 2 (bases 1 to 4342) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4342 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 99..230 /codon_start=1 /product="five tandem FLAG(R) epitope tags" /label=5xFLAG /translation="DYKDDDDKDYKDDDDKLMDYKDDDDKDYKDDDDKLMDYKDDDDK" terminator 257..444 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(312..336) /label=pHybLex-R /note="ADH terminator, reverse primer" gene 519..1875 /label=kanMX /note="yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)" promoter 519..862 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 863..1672 /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(930..949) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" terminator 1678..1875 /label=TEF terminator /note="Ashbya gossypii TEF terminator" primer_bind complement(1979..1998) /label=T7 /note="T7 promoter, forward primer" promoter complement(1980..1998) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2085..2102) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2256..2844) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2336..2355) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3015..3875) /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 3638..3657 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3876..3980) /gene="bla" /label=AmpR promoter primer_bind 4048..4066 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(4104..4126) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 4226..4245 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 4326..2 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 4326..1 /label=SP6 /note="SP6 promoter, forward primer" ORIGIN 1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggggaggtg 61 gaggtggagg tggaggtgga ggtttaatta atcatatgga ctacaaggac gacgatgaca 121 aggattacaa agatgacgac gataagctta tggactacaa ggacgacgat gacaaggatt 181 acaaagatga cgacgataag cttatggact acaaggacga cgatgacaag catatgggcg 241 cgccacttct aaataagcga atttcttatg atttatgatt tttattatta aataagttat 301 aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt 361 attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt atgaggtcgc 421 tcttattgac cacacctcta ccggcagatc cgctagggat aacagggtaa tatagatctg 481 tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc cagaataccc 541 tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc ccgtacattt 601 agcccataca tccccatgta taatcatttg catccataca ttttgatggc cgcacggcgc 661 gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca gggaaacgct cccctcacag 721 acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt taggatttgc 781 cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca 841 tcacatccga acataaacaa ccatgggtaa ggaaaagact cacgtttcga ggccgcgatt 901 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca 961 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa 1021 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct 1081 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg 1141 gttactcacc actgcgatcc ccggcaaaac agcattccag gtattagaag aatatcctga 1201 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc 1261 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg 1321 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt 1381 tgaacaagtc tggaaagaaa tgcataagct tttgccattc tcaccggatt cagtcgtcac 1441 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat 1501 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg 1561 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa 1621 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagtact 1681 gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta 1741 gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc 1801 tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg 1861 ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccagttta aacgagctcg 1921 aattcatcga tgatatcaga tccactagtg gcctatgcgg ccgcggatct gccggtctcc 1981 ctatagtgag tcgtattaat ttcgataagc caggttaacc tgcattaatg aatcggccaa 2041 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 2101 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 2161 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 2221 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 2281 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 2341 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 2401 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc 2461 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 2521 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 2581 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 2641 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca 2701 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 2761 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 2821 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 2881 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 2941 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 3001 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 3061 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 3121 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 3181 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 3241 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 3301 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 3361 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 3421 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 3481 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 3541 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 3601 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 3661 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 3721 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 3781 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 3841 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 3901 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 3961 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc 4021 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg 4081 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct 4141 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc 4201 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat 4261 atggacatat tgtcgttaga acgcggctac aattaataca taaccttatg tatcatacac 4321 atacgattta ggtgacacta ta //