LOCUS       sequence-72655-e        4342 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  genomic G11-5FLAG epitope tagging in S. pombe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4342)
  AUTHORS   Gadaleta MC, Iwasaki O, Noguchi C, Noma K, Noguchi E
  TITLE     New vectors for epitope tagging and gene disruption in 
            Schizosaccharomyces pombe.
  JOURNAL   Biotechniques. 2013 Nov;55(5):257-63. doi: 10.2144/000114100.
  PUBMED    24215641
REFERENCE   2  (bases 1 to 4342)
  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"; doi: "10"; journalName: 
            "Biotechniques"; date: "2013-11"; volume: "55"; issue: "5"; pages: 
            "257-63"
COMMENT     Sequence Label: pFA6a-G11-5FLAG-kanMX6
FEATURES             Location/Qualifiers
     source          1..4342
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             99..230
                     /codon_start=1
                     /product="five tandem FLAG(R) epitope tags"
                     /label=5xFLAG
                     /translation="DYKDDDDKDYKDDDDKLMDYKDDDDKDYKDDDDKLMDYKDDDDK"
     terminator      257..444
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(312..336)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            519..1875
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        519..862
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             863..1672
                     /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(930..949)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1678..1875
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1979..1998)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1980..1998)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2085..2102)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2256..2844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2336..2355)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3015..3806,3807..3875))
                     /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     3638..3657
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3876..3980)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4048..4066
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4104..4126)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4226..4245
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4326..4342,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4326..4342,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggggaggtg
       61 gaggtggagg tggaggtgga ggtttaatta atcatatgga ctacaaggac gacgatgaca
      121 aggattacaa agatgacgac gataagctta tggactacaa ggacgacgat gacaaggatt
      181 acaaagatga cgacgataag cttatggact acaaggacga cgatgacaag catatgggcg
      241 cgccacttct aaataagcga atttcttatg atttatgatt tttattatta aataagttat
      301 aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt
      361 attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt atgaggtcgc
      421 tcttattgac cacacctcta ccggcagatc cgctagggat aacagggtaa tatagatctg
      481 tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc cagaataccc
      541 tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc ccgtacattt
      601 agcccataca tccccatgta taatcatttg catccataca ttttgatggc cgcacggcgc
      661 gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca gggaaacgct cccctcacag
      721 acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt taggatttgc
      781 cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca
      841 tcacatccga acataaacaa ccatgggtaa ggaaaagact cacgtttcga ggccgcgatt
      901 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
      961 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
     1021 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     1081 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     1141 gttactcacc actgcgatcc ccggcaaaac agcattccag gtattagaag aatatcctga
     1201 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     1261 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     1321 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     1381 tgaacaagtc tggaaagaaa tgcataagct tttgccattc tcaccggatt cagtcgtcac
     1441 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     1501 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     1561 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     1621 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagtact
     1681 gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta
     1741 gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc
     1801 tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg
     1861 ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccagttta aacgagctcg
     1921 aattcatcga tgatatcaga tccactagtg gcctatgcgg ccgcggatct gccggtctcc
     1981 ctatagtgag tcgtattaat ttcgataagc caggttaacc tgcattaatg aatcggccaa
     2041 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2101 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2161 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2221 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2281 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2341 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2401 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     2461 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2521 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2581 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2641 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     2701 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2761 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2821 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2881 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2941 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3001 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3061 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3121 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3181 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3241 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3301 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3361 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3421 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3481 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3541 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3601 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3661 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3721 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3781 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3841 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3901 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3961 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     4021 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     4081 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     4141 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     4201 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     4261 atggacatat tgtcgttaga acgcggctac aattaataca taaccttatg tatcatacac
     4321 atacgattta ggtgacacta ta
//
