LOCUS       sequence-146281-        4112 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 4112)
  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 4112)
  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-HA3
FEATURES             Location/Qualifiers
     source          1..4112
                     /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             409..435
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             439..465
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             472..498
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      531..718
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(586..610)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            777..2133
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        777..1120
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1121..1930
                     /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(1188..1207)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1936..2133
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2192..2209)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2363..2951)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2443..2462)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3122..3913,3914..3982))
                     /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     3745..3764
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3983..4087)
                     /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 ctagcatgta cccatacgat
      421 gttcctgact atgcgggcta cccgtatgac gtgccggact atgcaggagc ctatccttat
      481 gacgttccag attacgctgg ttctagatga ggcgcgccac ttctaaataa gcgaatttct
      541 tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt
      601 aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg
      661 tcaggttgct ttctcaggta tagtatgagg tcgctcttat tgaccacacc tctaccggca
      721 gatccgctag gagatctgtt tagcttgcct cgtccccgcc gggtcacccg gccagcgaca
      781 tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg
      841 actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca tccatacatt
      901 ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcagacc tgcgagcagg
      961 gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg tagagaaata
     1021 taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt aaaatcttgc
     1081 taggatacag ttctcacatc acatccgaac ataaacaacc atgggtaagg aaaagactca
     1141 cgtttcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     1201 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc
     1261 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     1321 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     1381 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggcaaaacag cattccaggt
     1441 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     1501 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     1561 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     1621 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagcttt tgccattctc
     1681 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     1741 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     1801 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     1861 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     1921 gtttttctaa tcagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta
     1981 tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt
     2041 tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt aatatcatgc
     2101 gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta acgccgccat
     2161 ccagtttaaa cgagctcgaa ttctaggctt cctcgctcac tgactcgctg cgctcggtcg
     2221 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2281 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2341 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2401 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2461 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2521 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     2581 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2641 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2701 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2761 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     2821 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2881 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2941 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3001 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3061 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3121 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3181 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3241 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3301 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3361 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3421 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3481 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3541 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3601 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3661 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3721 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3781 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3841 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3901 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3961 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4021 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4081 ttccgcgcac atttccccga aaagtgccac ct
//
