LOCUS Exported 4789 bp ds-DNA circular SYN 24-12-2013 DEFINITION Plasmid with a kanMX marker for adding a C-terminal 12Pk tag. ACCESSION . VERSION . KEYWORDS pFA6a-12Pk-kanMX6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4789) 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;55:257-63. PUBMED 24215641 REFERENCE 2 (bases 1 to 4789) AUTHORS Noguchi Lab TITLE Direct Submission FEATURES Location/Qualifiers source 1..4789 /organism="synthetic DNA construct" /lab_host="Schizosaccharomyces pombe" /mol_type="other DNA" CDS 73..672 /codon_start=1 /product="12 tandem copies of the Pk (V5) epitope" /note="12Pk tag" /translation="GKPIPNPLLGLDSTSGGKPIPNPLLGLDSTLDGGGKPIPNPLLGL DSTSGGKPIPNPLLGLDSTLDGGGKPIPNPLLGLDSTSGGKPIPNPLLGLDSTLDGGGK PIPNPLLGLDSTSGGKPIPNPLLGLDSTLDGGGKPIPNPLLGLDSTSGGKPIPNPLLGL DSTLDGGGKPIPNPLLGLDSTSGGKPIPNPLLGLDST" terminator 702..889 /gene="S. cerevisiae ADH1" /note="ADH1 terminator" /note="transcription terminator for alcohol dehydrogenase 1" gene 964..2320 /note="kanMX" /note="yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)" promoter 964..1307 /note="TEF promoter" /note="Ashbya gossypii TEF promoter" CDS 1308..2117 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /note="KanR" /note="confers resistance to kanamycin" /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" terminator 2123..2320 /note="TEF terminator" /note="Ashbya gossypii TEF terminator" promoter complement(2425..2443) /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" rep_origin complement(2701..3289) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3460..4320) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(4321..4425) /gene="bla" /note="AmpR promoter" promoter 4771..4789 /note="SP6 promoter" /note="promoter for bacteriophage SP6 RNA polymerase" ORIGIN 1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac 61 ctcgacggtg gaggtaagcc tatccctaac cctctgctgg gtcttgattc tacatccgga 121 ggtaagccta tccctaaccc tctgctgggt cttgattcta cactcgacgg tggaggtaag 181 cctatcccta accctctgct gggtcttgat tctacatccg gaggtaagcc tatccctaac 241 cctctgctgg gtcttgattc tacactcgac ggtggaggta agcctatccc taaccctctg 301 ctgggtcttg attctacatc cggaggtaag cctatcccta accctctgct gggtcttgat 361 tctacactcg acggtggagg taagcctatc cctaaccctc tgctgggtct tgattctaca 421 tccggaggta agcctatccc taaccctctg ctgggtcttg attctacact cgacggtgga 481 ggtaagccta tccctaaccc tctgctgggt cttgattcta catccggagg taagcctatc 541 cctaaccctc tgctgggtct tgattctaca ctcgacggtg gaggtaagcc tatccctaac 601 cctctgctgg gtcttgattc tacatccgga ggtaagccta tccctaaccc tctgctgggt 661 cttgattcta cactcgagta gggcgcgcca cttctaaata agcgaatttc ttatgattta 721 tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact 781 cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc 841 tttctcaggt atagtatgag gtcgctctta ttgaccacac ctctaccggc agatccgcta 901 gggataacag ggtaatatag atctgtttag cttgcctcgt ccccgccggg tcacccggcc 961 agcgacatgg aggcccagaa taccctcctt gacagtcttg acgtgcgcag ctcaggggca 1021 tgatgtgact gtcgcccgta catttagccc atacatcccc atgtataatc atttgcatcc 1081 atacattttg atggccgcac ggcgcgaagc aaaaattacg gctcctcgct gcagacctgc 1141 gagcagggaa acgctcccct cacagacgcg ttgaattgtc cccacgccgc gcccctgtag 1201 agaaatataa aaggttagga tttgccactg aggttcttct ttcatatact tccttttaaa 1261 atcttgctag gatacagttc tcacatcaca tccgaacata aacaaccatg ggtaaggaaa 1321 agactcacgt ttcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata 1381 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc 1441 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag 1501 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt 1561 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggc aaaacagcat 1621 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt 1681 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat 1741 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg 1801 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aagcttttgc 1861 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg 1921 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc 1981 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc 2041 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc 2101 tcgatgagtt tttctaatca gtactgacaa taaaaagatt cttgttttca agaacttgtc 2161 atttgtatag tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta 2221 tatttttttt cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat 2281 atcatgcgtc aatcgtatgt gaatgctggt cgctatactg ctgtcgattc gatactaacg 2341 ccgccatcca gtttaaacga gctcgaattc atcgatgata tcagatccac tagtggccta 2401 tgcggccgcg gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt 2461 taacctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 2521 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 2581 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 2641 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 2701 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 2761 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 2821 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 2881 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 2941 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 3001 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 3061 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 3121 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 3181 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 3241 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 3301 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 3361 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 3421 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 3481 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 3541 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 3601 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 3661 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 3721 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 3781 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 3841 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 3901 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 3961 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 4021 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 4081 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 4141 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 4201 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 4261 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 4321 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 4381 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 4441 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 4501 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 4561 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 4621 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 4681 gcagattgta ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta 4741 atacataacc ttatgtatca tacacatacg atttaggtga cactataga //