LOCUS       sequence-146314-        4909 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 4909)
  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 4909)
  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-Myc13
FEATURES             Location/Qualifiers
     source          1..4909
                     /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             418..915
                     /codon_start=1
                     /product="13 tandem Myc epitope tags"
                     /label=13xMyc
                     /translation="EQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLIS
                     EEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLD
                     GEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISE
                     EDL"
     terminator      978..1165
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1033..1057)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             1730..2524
                     /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(2989..3006)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3160..3748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3240..3259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3919..4710,4711..4779))
                     /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     4542..4561
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4780..4884)
                     /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 ctagcgcgat taacggtgaa
      421 caaaagctaa tctccgagga agacttgaac ggtgaacaaa aattaatctc agaagaagac
      481 ttgaacggac tcgacggtga acaaaagttg atttctgaag aagatttgaa cggtgaacaa
      541 aagctaatct ccgaggaaga cttgaacggt gaacaaaaat taatctcaga agaagacttg
      601 aacggactcg acggtgaaca aaagttgatt tctgaagaag atttgaacgg tgaacaaaag
      661 ctaatctccg aggaagactt gaacggtgaa caaaaattaa tctcagaaga agacttgaac
      721 ggactcgacg gtgaacaaaa gttgatttct gaagaagatt tgaacggtga acaaaagcta
      781 atctccgagg aagacttgaa cggtgaacaa aaattaatct cagaagaaga cttgaacgga
      841 ctcgacggtg aacaaaagtt gatttctgaa gaagatttga acggtgaaca aaagctaatc
      901 tccgaggaag acttgaacgg tgaacaaaaa ttaatcaatc agtacaagtc tagatgaggc
      961 gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta
     1021 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct
     1081 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg
     1141 ctcttattga ccacacctct accggcagat ccgctaggag atctgagctc gatatcaagc
     1201 ttagctacaa atcccactgg ctatatgtat gcatttgtgt taaaaaagtt tgtatagatt
     1261 atttaatcta ctcagcattc tttctctaaa taggaatttg ttacttaatg gagaaaaaaa
     1321 tgtttcgatt tacctagtgt atttgtttgt atactcacgt ttaatttcaa acatccattc
     1381 tatcttgtgt aatttttggc atggtgaaaa agataatcag ccttataatc tttacaaaag
     1441 taagaaattc tgtaaataag ccttaatgcc cttgctttaa attaaaatgg ttctttttca
     1501 tgataatgtt tgcactttgt gaatatattt tagatagttc tgtgaggtat aattaagatg
     1561 ttttagagac ttatacaatt ttgtctttat aaattcttaa ttgattttac catcccagtt
     1621 taactatgct tcgtcggcat ctctgcacat gtcgtgtttt cttaccgtat tgtcctacca
     1681 agaacctctt ttttgcttgg atcgaaatta aaggtttaaa agcaaagtta tggatgctag
     1741 agtatttcaa agctattcag ctagagctga ggggatgaaa aatcccattg ccaaggaatt
     1801 gttggctttg atggaagaaa agcaaagcaa cttgtcagtc gcggtcgatt tgacgaagaa
     1861 atccgaaatc ttagaattgg tagataaaat tggaccctat gtctgtgtta tcaagacaca
     1921 tattgacgtt gtcgaggatt tcgaccagga tatggtagaa aaactggtgg ccttaggtaa
     1981 aaagcatcgt tttcttatct ttgaggatcg caaattcgca gacattggaa ataccgtcaa
     2041 gctacaatat gcatctggtg tgtacaaaat tgcttcttgg gctcatatca caaattgcca
     2101 tacagtgcca ggcgagggta ttatacaagg cctcaaagaa gttggtttac ctttgggacg
     2161 tggtctcttg cttttggctg aaatgtcttc caaaggctct ttggctactg gttcctacac
     2221 agagaaaacc ttagaatggt ttgagaagca taccgatttt tgctttggct ttatagctgg
     2281 tcgtcgattt cctaaccttc aaagcgacta cataactatg tcccctggta tcggcttgga
     2341 tgttaaagga gacgggctgg gacagcaata tcgtactcct gaagaagtga ttgtaaactg
     2401 cggtagcgat atcatcattg ttggtcgtgg agtctatgga gctggtcgta atcctgttgt
     2461 cgaagccaag agatatagag aagctggttg gaaggcatat cagcaaagac tttctcagca
     2521 ttaaaaaaag actaatgtaa aatttttttg gttggttatt gaaaaagtcg atgccttgtt
     2581 tgcgtttgtt ttcctaggcg ttttatgtca gaaggcattt agaattagta tacaagtact
     2641 ctttggtaaa attttatgta gcgactaaaa tattaactat tatagataaa caccttggga
     2701 ataaaaagta atttgctata gtaatttatt aaacatgctc ctacaacatt accacaatct
     2761 tttctcttgg attgacattg aataagaaaa gagtgaattt ttttagactt gtaatgataa
     2821 ctatgtacaa agccaatgaa agatgtatgt agatgaatgt aaaataccat gtagacaaac
     2881 aagataaaac ttggttataa acattggtgt tggaacagaa taaattagat gtcaaaaagt
     2941 ttcgtcaata tcacaagctt gtttaaacga gctcgaattc taggcttcct cgctcactga
     3001 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3061 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3121 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3181 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3241 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3301 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     3361 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3421 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3481 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3541 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3601 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3661 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3721 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3781 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3841 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3901 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3961 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4021 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4081 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4141 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4201 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4261 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4321 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4381 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4441 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4501 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4561 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4621 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4681 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4741 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4801 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4861 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacct
//
