LOCUS       sequence-160593-        4463 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 4463)
  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 4463)
  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: pNX3b2-HA3
FEATURES             Location/Qualifiers
     source          1..4463
                     /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"
     promoter        777..1120
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(2543..2560)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2714..3302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2794..2813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3473..4264,4265..4333))
                     /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     4096..4115
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4334..4438)
                     /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 gtcagcgaca
      781 tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg
      841 actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca tccatacatt
      901 ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcggacc tgcgagcagg
      961 gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg tagagaaata
     1021 taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt aaaatcttgc
     1081 taggatacag ttctcacatc acatccgaac ataaacaacc atgggtaaaa agcctgaact
     1141 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
     1201 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
     1261 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
     1321 ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag
     1381 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
     1441 cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga
     1501 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     1561 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     1621 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     1681 ggactgcccc gaagtccggc acctcatgca cgcggatttc ggctccaaca atgtcctgac
     1741 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     1801 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
     1861 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat
     1921 gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc
     1981 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
     2041 tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtagctag
     2101 ctaatcgatt agctgcagta atccttgctt aaaaagattc tcttttttta tgatatttgt
     2161 acataaactt tataaatgaa attcataata gaaacgacac gaaattacaa aatggaatat
     2221 gttcataggg tagacgaaac tatatacgca atctacatac atttatcaag aaggagaaaa
     2281 aggaggatgt aaaggaatac aggtaagcaa attgatacta atggctcaac gtgataagga
     2341 aaaagaattg cactttaaca ttaatattga caaggaggag ggcaccacac aaaaagttag
     2401 gtgtaacaga aaatcatgaa actatgattc ctaatttata tattggagga ttttctctaa
     2461 aaaaaaaaaa atacaacaaa taaaaaacac tcaatgacct gaccatttga tggagtttaa
     2521 acgagctcga attctaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2581 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2641 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2701 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2761 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2821 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2881 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     2941 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3001 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3061 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3121 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     3181 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3241 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3301 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3361 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3421 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3481 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3541 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3601 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3661 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3721 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3781 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3841 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3901 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3961 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4021 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4081 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4141 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4201 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4261 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4321 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4381 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4441 catttccccg aaaagtgcca cct
//
