LOCUS       sequence-72646-e        5611 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 5611)
  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 5611)
  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-LEU2MX6
FEATURES             Location/Qualifiers
     source          1..5611
                     /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"
     promoter        1183..1590
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             1591..2685
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(3246..3265)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3247..3265)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3352..3369)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3523..4111)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3603..3622)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4282..5073,5074..5142))
                     /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     4905..4924
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5143..5247)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5315..5333
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5371..5393)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5493..5512
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5593..5611
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5593..5610
                     /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 atcttgcagg catgcaagct tgtcgaggag aacttctagt
      961 atatctacat acctaatatt attgccttat taaaaatgga atcccaacaa ttacatcaaa
     1021 atccacattc tcttcaaaat caattgtcct gtacttcctt gttcatgtgt gttcaaaaac
     1081 gttatattta taggataatt atactctatt tctcaacaag taattggttg tttggccgag
     1141 cggtctaagg cgcctgattc aagaaatatc ttgaccgcag ttaactgtgg gaatactcag
     1201 gtatcgtaag atgcaagagt tcgaatctct tagcaaccat tatttttttc ctcaacataa
     1261 cgagaacaca caggggcgct atcgcacaga atcaaattcg atgactggaa attttttgtt
     1321 aatttcagag gtcgcctgac gcatatacct ttttcaactg aaaaattggg agaaaaagga
     1381 aaggtgagag ccgcggaacc ggcttttcat atagaataga gaagcgttca tgactaaatg
     1441 cttgcatcac aatacttgaa gttgacaata ttatttaagg acctattgtt ttttccaata
     1501 ggtggttagc aatcgtctta ctttctaact tttcttacct tttacatttc agcaatatat
     1561 atatatatat ttcaaggata taccattcta atgtctgccc ctaagaagat cgtcgttttg
     1621 ccaggtgacc acgttggtca agaaatcaca gccgaagcca ttaaggttct taaagctatt
     1681 tctgatgttc gttccaatgt caagttcgat ttcgaaaatc atttaattgg tggtgctgct
     1741 atcgatgcta caggtgtccc acttccagat gaggcgctgg aagcctccaa gaaggttgat
     1801 gccgttttgt taggtgctgt gggtggtcct aaatggggta ccggtagtgt tagacctgaa
     1861 caaggtttac taaaaatccg taaagaactt caattgtacg ccaacttaag accatgtaac
     1921 tttgcatccg actctctttt agacttatct ccaatcaagc cacaatttgc taaaggtact
     1981 gacttcgttg ttgtcagaga attagtggga ggtatttact ttggtaagag aaaggaagac
     2041 gatggtgatg gtgtcgcttg ggatagtgaa caatacaccg ttccagaagt gcaaagaatc
     2101 acaagaatgg ccgctttcat ggccctacaa catgagccac cattgcctat ttggtccttg
     2161 gataaagcta atgttttggc ctcttcaaga ttatggagaa aaactgtgga ggaaaccatc
     2221 aagaacgaat tccctacatt gaaggttcaa catcaattga ttgattctgc cgccatgatc
     2281 ctagttaaga acccaaccca cctaaatggt attataatca ccagcaacat gtttggtgat
     2341 atcatctccg atgaagcctc cgttatccca ggttccttgg gtttgttgcc atctgcgtcc
     2401 ttggcctctt tgccagacaa gaacaccgca tttggtttgt acgaaccatg ccacggttct
     2461 gctccagatt tgccaaagaa taaggtcaac cctatcgcca ctatcttgtc tgctgcaatg
     2521 atgttgaaat tgtcattgaa cttgcctgaa gaaggtaagg ccattgaaga tgcagttaaa
     2581 aaggttttgg atgcaggtat cagaactggt gatttaggtg gttccaacag taccacggaa
     2641 gtcggtgatg ctgtcgccga agaagttaag aaaatccttg cttaaaaaga ttctcttttt
     2701 ttatgatatt tgtacataaa ctttataaat gaaattcata atagaaacga cacgaaatta
     2761 caaaatggaa tatgttcata gggtagacga aactatatac gcaatctaca tacatttatc
     2821 aagaaggaga aaaaggagga tgtaaaggaa tacaggtaag caaattgata ctaatggctc
     2881 aacgtgataa ggaaaaagaa ttgcacttta acattaatat tgacaaggag gagggcacca
     2941 cacaaaaagt taggtgtaac agaaaatcat gaaactatga ttcctaattt atatattgga
     3001 ggattttctc taaaaaaaaa aaaatacaac aaataaaaaa cactcaatga cctgaccatt
     3061 tgatggagtt taagtcaata ccttcttgaa ccatttccca taatggtgaa agttccctca
     3121 agaattttac tctgtcagaa acggccttaa cgacgtagtc gatcgacaag ctgtttaaac
     3181 gagctcgaat tcatcgatga tatcagatcc actagtggcc tatgcggccg cggatctgcc
     3241 ggtctcccta tagtgagtcg tattaatttc gataagccag gttaacctgc attaatgaat
     3301 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3361 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3421 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3481 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3541 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3601 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3661 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     3721 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3781 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3841 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3901 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3961 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4021 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4081 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4141 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4201 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4261 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4321 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4381 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4441 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4501 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4561 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4621 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4681 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4741 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4801 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4861 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4921 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4981 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5041 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5101 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5161 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5221 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5281 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5341 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5401 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5461 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5521 gcaccatatg gacatattgt cgttagaacg cggctacaat taatacataa ccttatgtat
     5581 catacacata cgatttaggt gacactatag a
//
