LOCUS       sequence-146299-        4630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fission yeast epitope tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4630)
  AUTHORS   Amelina H, Moiseeva V, Collopy LC, Pearson SR, Armstrong CA, Tomita 
            K
  TITLE     Sequential and counter-selectable cassettes for fission yeast.
  JOURNAL   BMC Biotechnol. 2016 Nov 8;16(1):76.
  PUBMED    27825338
REFERENCE   2  (bases 1 to 4630)
  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"; journalName: "BMC 
            Biotechnol."; date: "2016-11-8"; volume: "16(1)"; pages: "76"
COMMENT     Sequence Label: pNX3d-PK6
FEATURES             Location/Qualifiers
     source          1..4630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..61
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(99..121)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     221..240
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        321..339
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     321..338
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             406..447
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             451..492
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             493..534
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             541..582
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             586..627
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             628..669
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     terminator      699..886
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(754..778)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             1451..2245
                     /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(2710..2727)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2881..3469)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2961..2980)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3640..4431,4432..4500))
                     /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     4263..4282
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4501..4605)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
       61 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
      121 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
      181 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
      241 ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc
      301 ttatgtatca tacacatacg atttaggtga cactatagaa cgcggccgcc agctgaagct
      361 tcgtacgctg caggtcgacg gatccccggg ttaattaacg ctagcggtaa gcctattcct
      421 aaccctcttt tgggtctcga ttctacaggt ggaaaaccaa tccccaaccc actcctcggc
      481 cttgactcaa ctggcaaacc cataccaaat cctttacttg gattagatag tacctctagc
      541 ggtaagccta ttcctaaccc tcttttgggt ctcgattcta caggtggaaa accaatcccc
      601 aacccactcc tcggccttga ctcaactggc aaacccatac caaatccttt acttggatta
      661 gatagtacct ctagatgagg cgcgccactt ctaaataagc gaatttctta tgatttatga
      721 tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt
      781 aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt
      841 ctcaggtata gtatgaggtc gctcttattg accacacctc taccggcaga tccgctagga
      901 gatctgagct cgatatcaag cttagctaca aatcccactg gctatatgta tgcatttgtg
      961 ttaaaaaagt ttgtatagat tatttaatct actcagcatt ctttctctaa ataggaattt
     1021 gttacttaat ggagaaaaaa atgtttcgat ttacctagtg tatttgtttg tatactcacg
     1081 tttaatttca aacatccatt ctatcttgtg taatttttgg catggtgaaa aagataatca
     1141 gccttataat ctttacaaaa gtaagaaatt ctgtaaataa gccttaatgc ccttgcttta
     1201 aattaaaatg gttctttttc atgataatgt ttgcactttg tgaatatatt ttagatagtt
     1261 ctgtgaggta taattaagat gttttagaga cttatacaat tttgtcttta taaattctta
     1321 attgatttta ccatcccagt ttaactatgc ttcgtcggca tctctgcaca tgtcgtgttt
     1381 tcttaccgta ttgtcctacc aagaacctct tttttgcttg gatcgaaatt aaaggtttaa
     1441 aagcaaagtt atggatgcta gagtatttca aagctattca gctagagctg aggggatgaa
     1501 aaatcccatt gccaaggaat tgttggcttt gatggaagaa aagcaaagca acttgtcagt
     1561 cgcggtcgat ttgacgaaga aatccgaaat cttagaattg gtagataaaa ttggacccta
     1621 tgtctgtgtt atcaagacac atattgacgt tgtcgaggat ttcgaccagg atatggtaga
     1681 aaaactggtg gccttaggta aaaagcatcg ttttcttatc tttgaggatc gcaaattcgc
     1741 agacattgga aataccgtca agctacaata tgcatctggt gtgtacaaaa ttgcttcttg
     1801 ggctcatatc acaaattgcc atacagtgcc aggcgagggt attatacaag gcctcaaaga
     1861 agttggttta cctttgggac gtggtctctt gcttttggct gaaatgtctt ccaaaggctc
     1921 tttggctact ggttcctaca cagagaaaac cttagaatgg tttgagaagc ataccgattt
     1981 ttgctttggc tttatagctg gtcgtcgatt tcctaacctt caaagcgact acataactat
     2041 gtcccctggt atcggcttgg atgttaaagg agacgggctg ggacagcaat atcgtactcc
     2101 tgaagaagtg attgtaaact gcggtagcga tatcatcatt gttggtcgtg gagtctatgg
     2161 agctggtcgt aatcctgttg tcgaagccaa gagatataga gaagctggtt ggaaggcata
     2221 tcagcaaaga ctttctcagc attaaaaaaa gactaatgta aaattttttt ggttggttat
     2281 tgaaaaagtc gatgccttgt ttgcgtttgt tttcctaggc gttttatgtc agaaggcatt
     2341 tagaattagt atacaagtac tctttggtaa aattttatgt agcgactaaa atattaacta
     2401 ttatagataa acaccttggg aataaaaagt aatttgctat agtaatttat taaacatgct
     2461 cctacaacat taccacaatc ttttctcttg gattgacatt gaataagaaa agagtgaatt
     2521 tttttagact tgtaatgata actatgtaca aagccaatga aagatgtatg tagatgaatg
     2581 taaaatacca tgtagacaaa caagataaaa cttggttata aacattggtg ttggaacaga
     2641 ataaattaga tgtcaaaaag tttcgtcaat atcacaagct tgtttaaacg agctcgaatt
     2701 ctaggcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2761 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2821 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2881 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2941 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3001 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3061 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3121 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3181 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3241 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3301 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     3361 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3421 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3481 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3541 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3601 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3661 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3721 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3781 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3841 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3901 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3961 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4021 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4081 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4141 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4201 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4261 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4321 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4381 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4441 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4501 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4561 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4621 agtgccacct
//
