LOCUS       sequence-72641-e        5008 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 5008)
  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 5008)
  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-hphMX6
FEATURES             Location/Qualifiers
     source          1..5008
                     /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..2539
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        964..1307
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1308..2336
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             1308..2336
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2342..2539
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2643..2662)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2644..2662)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2749..2766)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2920..3508)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3000..3019)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3679..4470,4471..4539))
                     /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     4302..4321
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4540..4644)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4712..4730
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4768..4790)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4890..4909
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4990..5008
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4990..5007
                     /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 ggtaaaaagc
     1321 ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg
     1381 acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc
     1441 gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt
     1501 atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca
     1561 gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc
     1621 ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg
     1681 cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat
     1741 acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa
     1801 ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt
     1861 gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg
     1921 tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg
     1981 attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc
     2041 agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg
     2101 cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg
     2161 atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg
     2221 tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag
     2281 tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaataatcag
     2341 tactgacaat aaaaagattc ttgttttcaa gaacttgtca tttgtatagt ttttttatat
     2401 tgtagttgtt ctattttaat caaatgttag cgtgatttat attttttttc gcctcgacat
     2461 catctgccca gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg
     2521 aatgctggtc gctatactgc tgtcgattcg atactaacgc cgccatccag tttaaacgag
     2581 ctcgaattca tcgatgatat cagatccact agtggcctat gcggccgcgg atctgccggt
     2641 ctccctatag tgagtcgtat taatttcgat aagccaggtt aacctgcatt aatgaatcgg
     2701 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2761 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2821 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2881 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2941 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3001 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3061 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     3121 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3181 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3241 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3301 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3361 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3421 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3481 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3541 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3601 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3661 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3721 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3781 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3841 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3901 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3961 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4021 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4081 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4141 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4201 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4261 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4321 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4381 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4441 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4501 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4561 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4621 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     4681 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     4741 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     4801 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     4861 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     4921 ccatatggac atattgtcgt tagaacgcgg ctacaattaa tacataacct tatgtatcat
     4981 acacatacga tttaggtgac actataga
//
