LOCUS       sequence-146282-        4559 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 4559)
  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 4559)
  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: pNX3a-Myc13
FEATURES             Location/Qualifiers
     source          1..4559
                     /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"
     gene            1224..2580
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1224..1567
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1568..2377
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1635..1654)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2383..2580
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2639..2656)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2810..3398)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2890..2909)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3569..4360,4361..4429))
                     /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     4192..4211
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4430..4534)
                     /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 atctgtttag cttgcctcgt
     1201 ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg
     1261 acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc
     1321 atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg
     1381 gctcctcgct gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc
     1441 cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct
     1501 ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata
     1561 aacaaccatg ggtaaggaaa agactcacgt ttcgaggccg cgattaaatt ccaacatgga
     1621 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     1681 ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     1741 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     1801 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     1861 gatccccggc aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     1921 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     1981 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2041 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2101 agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2161 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2221 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2281 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     2341 attgcagttt catttgatgc tcgatgagtt tttctaatca gtactgacaa taaaaagatt
     2401 cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa
     2461 tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag
     2521 ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg
     2581 ctgtcgattc gatactaacg ccgccatcca gtttaaacga gctcgaattc taggcttcct
     2641 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2701 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2761 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2821 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2881 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2941 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3001 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3061 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3121 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3181 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3241 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3301 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3361 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3421 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3481 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3541 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3601 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3661 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3721 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3781 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3841 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3901 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3961 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4021 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4081 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4141 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4201 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4261 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4321 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4381 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4441 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4501 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacct
//
