LOCUS       sequence-72648-e        5123 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 5123)
  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 5123)
  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-ura4MX6
FEATURES             Location/Qualifiers
     source          1..5123
                     /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"
     CDS             complement(1358..2152)
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe ura4"
                     /product="orotidine 5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=S. pombe ura4
                     /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
                     EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
                     LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
                     TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
                     NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
     primer_bind     complement(2758..2777)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2759..2777)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2864..2881)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3035..3623)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3115..3134)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3794..4585,4586..4654))
                     /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     4417..4436
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4655..4759)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4827..4845
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4883..4905)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5005..5024
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5105..5123
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5105..5122
                     /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 atctcttgtg atattgacga aactttttga catctaattt
      961 attctgttcc aacaccaatg tttataacca agttttatct tgtttgtcta catggtattt
     1021 tacattcatc tacatacatc tttcattggc tttgtacata gttatcatta caagtctaaa
     1081 aaaattcact cttttcttat tcaatgtcaa tccaagagaa aagattgtgg taatgttgta
     1141 ggagcatgtt taataaatta ctatagcaaa ttacttttta ttcccaaggt gtttatctat
     1201 aatagttaat attttagtcg ctacataaaa ttttaccaaa gagtacttgt atactaattc
     1261 taaatgcctt ctgacataaa acgcctagga aaacaaacgc aaacaaggca tcgacttttt
     1321 caataaccaa ccaaaaaaat tttacattag tcttttttta atgctgagaa agtctttgct
     1381 gatatgcctt ccaaccagct tctctatatc tcttggcttc gacaacagga ttacgaccag
     1441 ctccatagac tccacgacca acaatgatga tatcgctacc gcagtttaca atcacttctt
     1501 caggagtacg atattgctgt cccagcccgt ctcctttaac atccaagccg ataccagggg
     1561 acatagttat gtagtcgctt tgaaggttag gaaatcgacg accagctata aagccaaagc
     1621 aaaaatcggt atgcttctca aaccattcta aggttttctc tgtgtaggaa ccagtagcca
     1681 aagagccttt ggaagacatt tcagccaaaa gcaagagacc acgtcccaaa ggtaaaccaa
     1741 cttctttgag gccttgtata ataccctcgc ctggcactgt atggcaattt gtgatatgag
     1801 cccaagaagc aattttgtac acaccagatg catattgtag cttgacggta tttccaatgt
     1861 ctgcgaattt gcgatcctca aagataagaa aacgatgctt tttacctaag gccaccagtt
     1921 tttctaccat atcctggtcg aaatcctcga caacgtcaat atgtgtcttg ataacacaga
     1981 catagggtcc aattttatct accaattcta agatttcgga tttcttcgtc aaatcgaccg
     2041 cgactgacaa gttgctttgc ttttcttcca tcaaagccaa caattccttg gcaatgggat
     2101 ttttcatccc ctcagctcta gctgaatagc tttgaaatac tctagcatcc ataactttgc
     2161 ttttaaacct ttaatttcga tccaagcaaa aaagaggttc ttggtaggac aatacggtaa
     2221 gaaaacacga catgtgcaga gatgccgacg aagcatagtt aaactgggat ggtaaaatca
     2281 attaagaatt tataaagaca aaattgtata agtctctaaa acatcttaat tatacctcac
     2341 agaactatct aaaatatatt cacaaagtgc aaacattatc atgaaaaaga accattttaa
     2401 tttaaagcaa gggcattaag gcttatttac agaatttctt acttttgtaa agattataag
     2461 gctgattatc tttttcacca tgccaaaaat tacacaagat agaatggatg tttgaaatta
     2521 aacgtgagta tacaaacaaa tacactaggt aaatcgaaac atttttttct ccattaagta
     2581 acaaattcct atttagagaa agaatgctga gtagattaaa taatctatac aaactttttt
     2641 aacacaaatg catacatata gccagtggga tttgtagcta agctgtttaa acgagctcga
     2701 attcatcgat gatatcagat ccactagtgg cctatgcggc cgcggatctg ccggtctccc
     2761 tatagtgagt cgtattaatt tcgataagcc aggttaacct gcattaatga atcggccaac
     2821 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2881 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2941 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3001 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3061 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3121 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3181 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     3241 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3301 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3361 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3421 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     3481 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3541 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3601 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3661 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3721 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3781 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3841 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3901 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3961 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4021 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4081 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4141 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4201 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4261 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4321 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4381 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4441 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4501 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4561 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4621 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4681 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4741 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     4801 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     4861 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     4921 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     4981 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     5041 tggacatatt gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca
     5101 tacgatttag gtgacactat aga
//
