LOCUS       sequence-146300-        4765 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 4765)
  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 4765)
  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-PK9
FEATURES             Location/Qualifiers
     source          1..4765
                     /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"
     terminator      834..1021
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(889..913)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             1586..2380
                     /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(2845..2862)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3016..3604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3096..3115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3775..4566,4567..4635))
                     /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     4398..4417
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4636..4740)
                     /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 tacctctaga tgaggcgcgc cacttctaaa taagcgaatt
      841 tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat
      901 tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt
      961 aggtcaggtt gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg
     1021 gcagatccgc taggagatct gagctcgata tcaagcttag ctacaaatcc cactggctat
     1081 atgtatgcat ttgtgttaaa aaagtttgta tagattattt aatctactca gcattctttc
     1141 tctaaatagg aatttgttac ttaatggaga aaaaaatgtt tcgatttacc tagtgtattt
     1201 gtttgtatac tcacgtttaa tttcaaacat ccattctatc ttgtgtaatt tttggcatgg
     1261 tgaaaaagat aatcagcctt ataatcttta caaaagtaag aaattctgta aataagcctt
     1321 aatgcccttg ctttaaatta aaatggttct ttttcatgat aatgtttgca ctttgtgaat
     1381 atattttaga tagttctgtg aggtataatt aagatgtttt agagacttat acaattttgt
     1441 ctttataaat tcttaattga ttttaccatc ccagtttaac tatgcttcgt cggcatctct
     1501 gcacatgtcg tgttttctta ccgtattgtc ctaccaagaa cctctttttt gcttggatcg
     1561 aaattaaagg tttaaaagca aagttatgga tgctagagta tttcaaagct attcagctag
     1621 agctgagggg atgaaaaatc ccattgccaa ggaattgttg gctttgatgg aagaaaagca
     1681 aagcaacttg tcagtcgcgg tcgatttgac gaagaaatcc gaaatcttag aattggtaga
     1741 taaaattgga ccctatgtct gtgttatcaa gacacatatt gacgttgtcg aggatttcga
     1801 ccaggatatg gtagaaaaac tggtggcctt aggtaaaaag catcgttttc ttatctttga
     1861 ggatcgcaaa ttcgcagaca ttggaaatac cgtcaagcta caatatgcat ctggtgtgta
     1921 caaaattgct tcttgggctc atatcacaaa ttgccataca gtgccaggcg agggtattat
     1981 acaaggcctc aaagaagttg gtttaccttt gggacgtggt ctcttgcttt tggctgaaat
     2041 gtcttccaaa ggctctttgg ctactggttc ctacacagag aaaaccttag aatggtttga
     2101 gaagcatacc gatttttgct ttggctttat agctggtcgt cgatttccta accttcaaag
     2161 cgactacata actatgtccc ctggtatcgg cttggatgtt aaaggagacg ggctgggaca
     2221 gcaatatcgt actcctgaag aagtgattgt aaactgcggt agcgatatca tcattgttgg
     2281 tcgtggagtc tatggagctg gtcgtaatcc tgttgtcgaa gccaagagat atagagaagc
     2341 tggttggaag gcatatcagc aaagactttc tcagcattaa aaaaagacta atgtaaaatt
     2401 tttttggttg gttattgaaa aagtcgatgc cttgtttgcg tttgttttcc taggcgtttt
     2461 atgtcagaag gcatttagaa ttagtataca agtactcttt ggtaaaattt tatgtagcga
     2521 ctaaaatatt aactattata gataaacacc ttgggaataa aaagtaattt gctatagtaa
     2581 tttattaaac atgctcctac aacattacca caatcttttc tcttggattg acattgaata
     2641 agaaaagagt gaattttttt agacttgtaa tgataactat gtacaaagcc aatgaaagat
     2701 gtatgtagat gaatgtaaaa taccatgtag acaaacaaga taaaacttgg ttataaacat
     2761 tggtgttgga acagaataaa ttagatgtca aaaagtttcg tcaatatcac aagcttgttt
     2821 aaacgagctc gaattctagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2881 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2941 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3001 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3061 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3121 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3181 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     3241 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3301 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3361 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3421 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     3481 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3541 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3601 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3661 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3721 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3781 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3841 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3901 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3961 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4021 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4081 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4141 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4201 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4261 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4321 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4381 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4441 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4501 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4561 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4621 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4681 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4741 cacatttccc cgaaaagtgc cacct
//
