LOCUS       sequence-146304-        4462 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 4462)
  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 4462)
  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-HA3
FEATURES             Location/Qualifiers
     source          1..4462
                     /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"
     CDS             1283..2077
                     /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(2542..2559)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2713..3301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2793..2812)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3472..4263,4264..4332))
                     /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     4095..4114
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4333..4437)
                     /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 gagatctgag ctcgatatca agcttagcta caaatcccac tggctatatg
      781 tatgcatttg tgttaaaaaa gtttgtatag attatttaat ctactcagca ttctttctct
      841 aaataggaat ttgttactta atggagaaaa aaatgtttcg atttacctag tgtatttgtt
      901 tgtatactca cgtttaattt caaacatcca ttctatcttg tgtaattttt ggcatggtga
      961 aaaagataat cagccttata atctttacaa aagtaagaaa ttctgtaaat aagccttaat
     1021 gcccttgctt taaattaaaa tggttctttt tcatgataat gtttgcactt tgtgaatata
     1081 ttttagatag ttctgtgagg tataattaag atgttttaga gacttataca attttgtctt
     1141 tataaattct taattgattt taccatccca gtttaactat gcttcgtcgg catctctgca
     1201 catgtcgtgt tttcttaccg tattgtccta ccaagaacct cttttttgct tggatcgaaa
     1261 ttaaaggttt aaaagcaaag ttatggatgc tagagtattt caaagctatt cagctagagc
     1321 tgaggggatg aaaaatccca ttgccaagga attgttggct ttgatggaag aaaagcaaag
     1381 caacttgtca gtcgcggtcg atttgacgaa gaaatccgaa atcttagaat tggtagataa
     1441 aattggaccc tatgtctgtg ttatcaagac acatattgac gttgtcgagg atttcgacca
     1501 ggatatggta gaaaaactgg tggccttagg taaaaagcat cgttttctta tctttgagga
     1561 tcgcaaattc gcagacattg gaaataccgt caagctacaa tatgcatctg gtgtgtacaa
     1621 aattgcttct tgggctcata tcacaaattg ccatacagtg ccaggcgagg gtattataca
     1681 aggcctcaaa gaagttggtt tacctttggg acgtggtctc ttgcttttgg ctgaaatgtc
     1741 ttccaaaggc tctttggcta ctggttccta cacagagaaa accttagaat ggtttgagaa
     1801 gcataccgat ttttgctttg gctttatagc tggtcgtcga tttcctaacc ttcaaagcga
     1861 ctacataact atgtcccctg gtatcggctt ggatgttaaa ggagacgggc tgggacagca
     1921 atatcgtact cctgaagaag tgattgtaaa ctgcggtagc gatatcatca ttgttggtcg
     1981 tggagtctat ggagctggtc gtaatcctgt tgtcgaagcc aagagatata gagaagctgg
     2041 ttggaaggca tatcagcaaa gactttctca gcattaaaaa aagactaatg taaaattttt
     2101 ttggttggtt attgaaaaag tcgatgcctt gtttgcgttt gttttcctag gcgttttatg
     2161 tcagaaggca tttagaatta gtatacaagt actctttggt aaaattttat gtagcgacta
     2221 aaatattaac tattatagat aaacaccttg ggaataaaaa gtaatttgct atagtaattt
     2281 attaaacatg ctcctacaac attaccacaa tcttttctct tggattgaca ttgaataaga
     2341 aaagagtgaa tttttttaga cttgtaatga taactatgta caaagccaat gaaagatgta
     2401 tgtagatgaa tgtaaaatac catgtagaca aacaagataa aacttggtta taaacattgg
     2461 tgttggaaca gaataaatta gatgtcaaaa agtttcgtca atatcacaag cttgtttaaa
     2521 cgagctcgaa ttctaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2581 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2641 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2701 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2761 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2821 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2881 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     2941 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3001 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3061 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3121 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     3181 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3241 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3301 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3361 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3421 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3481 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3541 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3601 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3661 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3721 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3781 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3841 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3901 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3961 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4021 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4081 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4141 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4201 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4261 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4321 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4381 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4441 atttccccga aaagtgccac ct
//
