LOCUS       sequence-160589-        4775 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 4775)
  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 4775)
  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: pNX3b-Myc13
FEATURES             Location/Qualifiers
     source          1..4775
                     /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..2796
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        1224..1567
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1568..2593
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLMHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2599..2796
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2855..2872)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3026..3614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3106..3125)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3785..4576,4577..4645))
                     /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     4408..4427
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4646..4750)
                     /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 tcacccggtc agcgacatgg aggcccagaa taccctcctt gacagtcttg
     1261 acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc
     1321 atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg
     1381 gctcctcgct gcggacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc
     1441 cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct
     1501 ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata
     1561 aacaaccatg ggtaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     1621 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     1681 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     1741 ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt
     1801 gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     1861 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     1921 ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg
     1981 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     2041 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     2101 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcatgcacgc
     2161 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     2221 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     2281 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     2341 aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     2401 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     2461 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg
     2521 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     2581 gagggcaaag gaatcagtac tgacaataaa aagattcttg ttttcaagaa cttgtcattt
     2641 gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt
     2701 ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca
     2761 tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata ctaacgccgc
     2821 catccagttt aaacgagctc gaattctagg cttcctcgct cactgactcg ctgcgctcgg
     2881 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2941 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3001 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3061 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3121 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3181 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     3241 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3301 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3361 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3421 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     3481 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3541 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3601 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3661 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3721 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3781 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3841 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3901 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3961 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4021 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4081 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4141 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4201 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4261 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4321 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4381 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4441 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4501 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4561 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4621 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4681 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4741 gggttccgcg cacatttccc cgaaaagtgc cacct
//
