LOCUS       sequence-173240-        4280 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 4280)
  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 4280)
  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: pNX3a-PK9
FEATURES             Location/Qualifiers
     source          1..4280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            1..1357
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1..344
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             345..1154
                     /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(412..431)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1160..1357
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1416..1433)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1587..2175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1667..1686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2346..3137,3138..3206))
                     /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     2969..2988
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3207..3311)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3379..3397
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3435..3457)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3557..3576
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3657..3675
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3657..3674
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             3742..3783
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3787..3828
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3829..3870
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3877..3918
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3922..3963
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3964..4005
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     terminator      4035..4222
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4090..4114)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
ORIGIN
        1 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga
       61 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata
      121 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag
      181 cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga
      241 aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc
      301 ttgctaggat acagttctca catcacatcc gaacataaac aaccatgggt aaggaaaaga
      361 ctcacgtttc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
      421 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg
      481 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
      541 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
      601 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggcaaa acagcattcc
      661 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
      721 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
      781 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
      841 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat
      901 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
      961 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     1021 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     1081 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     1141 atgagttttt ctaatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt
     1201 tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat
     1261 tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc
     1321 atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg
     1381 ccatccagtt taaacgagct cgaattctag gcttcctcgc tcactgactc gctgcgctcg
     1441 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     1501 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1561 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1621 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1681 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1741 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1801 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1861 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1921 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1981 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2041 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2101 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2161 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2221 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2281 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2341 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2401 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     2461 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     2521 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2581 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     2641 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2701 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2761 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     2821 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     2881 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     2941 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3001 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3061 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3121 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3181 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3241 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3301 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     3361 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     3421 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     3481 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     3541 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatggacata
     3601 ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca catacgattt
     3661 aggtgacact atagaacgcg gccgccagct gaagcttcgt acgctgcagg tcgacggatc
     3721 cccgggttaa ttaacgctag cggtaagcct attcctaacc ctcttttggg tctcgattct
     3781 acaggtggaa aaccaatccc caacccactc ctcggccttg actcaactgg caaacccata
     3841 ccaaatcctt tacttggatt agatagtacc tctagcggta agcctattcc taaccctctt
     3901 ttgggtctcg attctacagg tggaaaacca atccccaacc cactcctcgg ccttgactca
     3961 actggcaaac ccataccaaa tcctttactt ggattagata gtacctctag atgaggcgcg
     4021 ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
     4081 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
     4141 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc
     4201 ttattgacca cacctctacc ggcagatccg ctaggagatc tgtttagctt gcctcgtccc
     4261 cgccgggtca cccggccagc
//
