LOCUS       sequence-72634-e        4789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  genomic 12Pk epitope tagging in S. pombe.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Nov;55(5):257-63. doi: 10.2144/000114100.
  PUBMED    24215641
REFERENCE   2  (bases 1 to 4789)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Biotechniques"; date: "2013-11"; volume: "55"; issue: "5"; pages: 
            "257-63"
COMMENT     Sequence Label: pFA6a-12Pk-kanMX6
FEATURES             Location/Qualifiers
     source          1..4789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             73..672
                     /codon_start=1
                     /product="12 tandem copies of the Pk (V5) epitope"
                     /label=12Pk tag
                     /translation="GKPIPNPLLGLDSTSGGKPIPNPLLGLDSTLDGGGKPIPNPLLGL
                     DSTSGGKPIPNPLLGLDSTLDGGGKPIPNPLLGLDSTSGGKPIPNPLLGLDSTLDGGGK
                     PIPNPLLGLDSTSGGKPIPNPLLGLDSTLDGGGKPIPNPLLGLDSTSGGKPIPNPLLGL
                     DSTLDGGGKPIPNPLLGLDSTSGGKPIPNPLLGLDST"
     terminator      702..889
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(757..781)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            964..2320
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        964..1307
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1308..2117
                     /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(1375..1394)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2123..2320
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2424..2443)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2425..2443)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2530..2547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2701..3289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2781..2800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3460..4251,4252..4320))
                     /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     4083..4102
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4321..4425)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4493..4511
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4549..4571)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4671..4690
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4771..4789
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4771..4788
                     /label=SP6
                     /note="SP6 promoter, forward primer"
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
//
