LOCUS       sequence-160574-        4900 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 4900)
  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 4900)
  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-PK12
FEATURES             Location/Qualifiers
     source          1..4900
                     /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"
     CDS             676..717
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             721..762
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             763..804
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             811..852
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             856..897
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             898..939
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     terminator      969..1156
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1024..1048)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             1721..2515
                     /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(2980..2997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3151..3739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3231..3250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3910..4701,4702..4770))
                     /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     4533..4552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4771..4875)
                     /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 ctagcggtaa gcctattcct aaccctcttt tgggtctcga ttctacaggt
      721 ggaaaaccaa tccccaaccc actcctcggc cttgactcaa ctggcaaacc cataccaaat
      781 cctttacttg gattagatag tacctctagc ggtaagccta ttcctaaccc tcttttgggt
      841 ctcgattcta caggtggaaa accaatcccc aacccactcc tcggccttga ctcaactggc
      901 aaacccatac caaatccttt acttggatta gatagtacct ctagatgagg cgcgccactt
      961 ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa
     1021 taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga
     1081 gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc gctcttattg
     1141 accacacctc taccggcaga tccgctagga gatctgagct cgatatcaag cttagctaca
     1201 aatcccactg gctatatgta tgcatttgtg ttaaaaaagt ttgtatagat tatttaatct
     1261 actcagcatt ctttctctaa ataggaattt gttacttaat ggagaaaaaa atgtttcgat
     1321 ttacctagtg tatttgtttg tatactcacg tttaatttca aacatccatt ctatcttgtg
     1381 taatttttgg catggtgaaa aagataatca gccttataat ctttacaaaa gtaagaaatt
     1441 ctgtaaataa gccttaatgc ccttgcttta aattaaaatg gttctttttc atgataatgt
     1501 ttgcactttg tgaatatatt ttagatagtt ctgtgaggta taattaagat gttttagaga
     1561 cttatacaat tttgtcttta taaattctta attgatttta ccatcccagt ttaactatgc
     1621 ttcgtcggca tctctgcaca tgtcgtgttt tcttaccgta ttgtcctacc aagaacctct
     1681 tttttgcttg gatcgaaatt aaaggtttaa aagcaaagtt atggatgcta gagtatttca
     1741 aagctattca gctagagctg aggggatgaa aaatcccatt gccaaggaat tgttggcttt
     1801 gatggaagaa aagcaaagca acttgtcagt cgcggtcgat ttgacgaaga aatccgaaat
     1861 cttagaattg gtagataaaa ttggacccta tgtctgtgtt atcaagacac atattgacgt
     1921 tgtcgaggat ttcgaccagg atatggtaga aaaactggtg gccttaggta aaaagcatcg
     1981 ttttcttatc tttgaggatc gcaaattcgc agacattgga aataccgtca agctacaata
     2041 tgcatctggt gtgtacaaaa ttgcttcttg ggctcatatc acaaattgcc atacagtgcc
     2101 aggcgagggt attatacaag gcctcaaaga agttggttta cctttgggac gtggtctctt
     2161 gcttttggct gaaatgtctt ccaaaggctc tttggctact ggttcctaca cagagaaaac
     2221 cttagaatgg tttgagaagc ataccgattt ttgctttggc tttatagctg gtcgtcgatt
     2281 tcctaacctt caaagcgact acataactat gtcccctggt atcggcttgg atgttaaagg
     2341 agacgggctg ggacagcaat atcgtactcc tgaagaagtg attgtaaact gcggtagcga
     2401 tatcatcatt gttggtcgtg gagtctatgg agctggtcgt aatcctgttg tcgaagccaa
     2461 gagatataga gaagctggtt ggaaggcata tcagcaaaga ctttctcagc attaaaaaaa
     2521 gactaatgta aaattttttt ggttggttat tgaaaaagtc gatgccttgt ttgcgtttgt
     2581 tttcctaggc gttttatgtc agaaggcatt tagaattagt atacaagtac tctttggtaa
     2641 aattttatgt agcgactaaa atattaacta ttatagataa acaccttggg aataaaaagt
     2701 aatttgctat agtaatttat taaacatgct cctacaacat taccacaatc ttttctcttg
     2761 gattgacatt gaataagaaa agagtgaatt tttttagact tgtaatgata actatgtaca
     2821 aagccaatga aagatgtatg tagatgaatg taaaatacca tgtagacaaa caagataaaa
     2881 cttggttata aacattggtg ttggaacaga ataaattaga tgtcaaaaag tttcgtcaat
     2941 atcacaagct tgtttaaacg agctcgaatt ctaggcttcc tcgctcactg actcgctgcg
     3001 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3061 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3121 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3181 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3241 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3301 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     3361 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3421 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3481 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3541 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     3601 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3661 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3721 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3781 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3841 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3901 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3961 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4021 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4081 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4141 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4201 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4261 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4321 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4381 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4441 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4501 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4561 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4621 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4681 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4741 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4801 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4861 aataggggtt ccgcgcacat ttccccgaaa agtgccacct
//
