LOCUS       sequence-149965-        3912 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 3912)
  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 3912)
  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: pNX3c-PK3
FEATURES             Location/Qualifiers
     source          1..3912
                     /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             406..447
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             451..492
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             493..534
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     terminator      567..754
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(622..646)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            813..1932
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        813..1156
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1157..1729
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1735..1932
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1992..2009)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2163..2751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2243..2262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2922..3713,3714..3782))
                     /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     3545..3564
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3783..3887)
                     /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 ctagcggtaa gcctattcct
      421 aaccctcttt tgggtctcga ttctacaggt ggaaaaccaa tccccaaccc actcctcggc
      481 cttgactcaa ctggcaaacc cataccaaat cctttacttg gattagatag tacctctaga
      541 gattgaggcg cgccacttct aaataagcga atttcttatg atttatgatt tttattatta
      601 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
      661 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt
      721 atgaggtcgc tcttattgac cacacctcta ccggcagatc cgctaggaga tctgtttagc
      781 ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat accctccttg
      841 acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtat atttagccca
      901 cacatcccca tgtataatca tttgcatcca tacattttga tggctgcacg gcgcgaagca
      961 aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc acagacgcgt
     1021 tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga
     1081 ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat
     1141 ccgaacataa acaaccatgg gtaccactct tgacgacacg gcttaccggt accgcaccag
     1201 tgtcccgggg gacgccgagg ccatcgaggc actggatggg tccttcacca ccgacaccgt
     1261 cttccgcgtc accgccaccg gggacggctt caccctgcgg gaggtgccgg tggacccgcc
     1321 cctgaccaag gtgttccccg acgacgaatc ggacgacgaa tcggacgacg gggaggacgg
     1381 cgacccggac tcccggacgt tcgtcgcgta cggggacgac ggcgacctgg cgggcttcgt
     1441 ggtcgtctcg tactccggct ggaaccgccg gctgaccgtc gaggacatcg aggtcgcccc
     1501 ggagcaccgg gggcacgggg tcgggcgcgc gttgatgggg ctcgcgacgg agttcgcccg
     1561 cgagcggggc gccgggcacc tctggctgga ggtcaccaac gtcaacgcac cggcgatcca
     1621 cgcgtaccgg cggatggggt tcaccctctg cggcctggac accgccctgt acgacggcac
     1681 cgcctcggac ggcgagcagg cgctctacat gagcatgccc tgcccctaat cagtactgac
     1741 aataaaaaga ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt
     1801 gttctatttt aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc
     1861 ccagatgcga agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg
     1921 gtcgctatac tgctgtcgat tcgatactaa cgccgccatc cagtgttaaa cgagctcgaa
     1981 ttctaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2041 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2101 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2161 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2221 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2281 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2341 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2401 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2461 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2521 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2581 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     2641 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2701 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2761 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2821 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2881 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2941 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3001 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3061 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3121 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3181 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3241 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3301 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3361 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3421 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3481 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3541 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3601 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3661 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3721 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3781 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3841 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3901 aaagtgccac ct
//
