LOCUS       sequence-146374-        3998 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 3998)
  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 3998)
  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: pNX3s2-HA3
FEATURES             Location/Qualifiers
     source          1..3998
                     /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"
     polyA_signal    738..819
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     763..782
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(833..1207)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVCVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(1226..1273)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(1370..1394)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1391..1594)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1420..1440)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1595..1898
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(1972..1991)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1973..1991)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2078..2095)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2249..2837)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2329..2348)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3008..3799,3800..3868))
                     /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     3631..3650
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3869..3973)
                     /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 gagatctgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
      781 gctttatttg taaccattat aagctgcaat aaacaagttt cgaggtcgag tgtcagtcct
      841 gctcctcggc cacgaagtgc acgcagttgc cggccgggtc gcgcagggcg aactcccgcc
      901 cccacggctg ctcgccgatc tcggtcatgg ccggcccgga ggcgtcccgg aagttcgtgg
      961 acacgacctc cgaccactcg gcgtacagct cgtccaggcc gcgcacacac acccaggcca
     1021 gggtgttgtc cggcaccacc tggtcctgga ccgcgctgat gaacagggtc acgtcgtccc
     1081 ggaccacacc ggcgaagtcg tcctccacga agtcccggga gaacccgagc cggtcggtcc
     1141 agaactcgac cgctccggcg acgtcgcgcg cggtgagcac cggaacggca ctggtcaact
     1201 tggccatggt ttagttcctc accttgtcgt attatactat gccgatatac tatgccgatg
     1261 attaattgtc aacacggtcc gttccaatgc accgttcccg gccgcggagg ctggatcggt
     1321 cccggtgtct tctatggagg tcaaaacagc gtggatggcg tctccaggcg atctgacggt
     1381 tcactaaacg agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg
     1441 tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca
     1501 aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc
     1561 cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt
     1621 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     1681 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     1741 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     1801 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     1861 gtcagccagg cgggccattt accgtaagtt atgtaacggt ttaaacgagc tcgaattcat
     1921 cgatgatatc agatccacta gtggcctatg cggccgcgga tctgccggtc tccctatagt
     1981 gagtcgtatt aatttcgata agccaggtta acctgcatta atgaatcggc caacgcgcgg
     2041 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2101 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2161 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2221 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2281 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2341 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2401 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
     2461 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2521 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2581 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2641 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     2701 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2761 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2821 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2881 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2941 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3001 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3061 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3121 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3181 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3241 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3301 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3361 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3421 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3481 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3541 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3601 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3661 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3721 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3781 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3841 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3901 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3961 taggggttcc gcgcacattt ccccgaaaag tgccacct
//
