LOCUS       sequence-72650-e        4336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  genomic G9-5FLAG epitope tagging in S. pombe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4336)
  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 4336)
  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-G9-5FLAG-kanMX6
FEATURES             Location/Qualifiers
     source          1..4336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             93..224
                     /codon_start=1
                     /product="five tandem FLAG(R) epitope tags"
                     /label=5xFLAG
                     /translation="DYKDDDDKDYKDDDDKLMDYKDDDDKDYKDDDDKLMDYKDDDDK"
     terminator      251..438
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(306..330)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            513..1869
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        513..856
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             857..1666
                     /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(924..943)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1672..1869
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1973..1992)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1974..1992)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2079..2096)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2250..2838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2330..2349)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3009..3800,3801..3869))
                     /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     3632..3651
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3870..3974)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4042..4060
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4098..4120)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4220..4239
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4320..4336,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4320..4336,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggggaggtg
       61 gaggtggagg tggaggttta attaatcata tggactacaa ggacgacgat gacaaggatt
      121 acaaagatga cgacgataag cttatggact acaaggacga cgatgacaag gattacaaag
      181 atgacgacga taagcttatg gactacaagg acgacgatga caagcatatg ggcgcgccac
      241 ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa
      301 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt
      361 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg tcgctcttat
      421 tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga tctgtttagc
      481 ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat accctccttg
      541 acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac atttagccca
      601 tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg gcgcgaagca
      661 aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc acagacgcgt
      721 tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga
      781 ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat
      841 ccgaacataa acaaccatgg gtaaggaaaa gactcacgtt tcgaggccgc gattaaattc
      901 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
      961 tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     1021 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     1081 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     1141 caccactgcg atccccggca aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     1201 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     1261 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     1321 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     1381 agtctggaaa gaaatgcata agcttttgcc attctcaccg gattcagtcg tcactcatgg
     1441 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     1501 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     1561 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     1621 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag tactgacaat
     1681 aaaaagattc ttgttttcaa gaacttgtca tttgtatagt ttttttatat tgtagttgtt
     1741 ctattttaat caaatgttag cgtgatttat attttttttc gcctcgacat catctgccca
     1801 gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg aatgctggtc
     1861 gctatactgc tgtcgattcg atactaacgc cgccatccag tttaaacgag ctcgaattca
     1921 tcgatgatat cagatccact agtggcctat gcggccgcgg atctgccggt ctccctatag
     1981 tgagtcgtat taatttcgat aagccaggtt aacctgcatt aatgaatcgg ccaacgcgcg
     2041 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2101 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2161 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2221 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2281 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2341 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2401 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     2461 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2521 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2581 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2641 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     2701 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2761 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2821 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2881 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2941 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3001 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3061 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3121 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3181 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3241 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3301 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3361 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3421 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3481 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3541 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3601 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3661 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3721 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3781 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3841 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3901 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3961 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     4021 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     4081 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     4141 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     4201 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatggac
     4261 atattgtcgt tagaacgcgg ctacaattaa tacataacct tatgtatcat acacatacga
     4321 tttaggtgac actata
//
