LOCUS       sequence-160590-        4181 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 4181)
  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 4181)
  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-PK9
FEATURES             Location/Qualifiers
     source          1..4181
                     /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"
     CDS             541..582
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             586..627
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             628..669
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             676..717
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             721..762
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             763..804
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     terminator      837..1024
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(892..916)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1083..2202
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        1083..1426
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1427..1999
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      2005..2202
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2261..2278)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2432..3020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2512..2531)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3191..3982,3983..4051))
                     /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     3814..3833
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4052..4156)
                     /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 tacctctagc
      541 ggtaagccta ttcctaaccc tcttttgggt ctcgattcta caggtggaaa accaatcccc
      601 aacccactcc tcggccttga ctcaactggc aaacccatac caaatccttt acttggatta
      661 gatagtacct ctagcggtaa gcctattcct aaccctcttt tgggtctcga ttctacaggt
      721 ggaaaaccaa tccccaaccc actcctcggc cttgactcaa ctggcaaacc cataccaaat
      781 cctttacttg gattagatag tacctctaga gattgaggcg cgccacttct aaataagcga
      841 atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca
      901 aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc
      961 tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta
     1021 ccggcagatc cgctaggaga tctgtttagc ttgcctcgtc cccgccgggt cacccggcca
     1081 gcgacatgga ggcccagaat accctccttg acagtcttga cgtgcgcagc tcaggggcat
     1141 gatgtgactg tcgcccgtat atttagccca cacatcccca tgtataatca tttgcatcca
     1201 tacattttga tggctgcacg gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg
     1261 agcagggaaa cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga
     1321 gaaatataaa aggttaggat ttgccactga ggttcttctt tcatatactt ccttttaaaa
     1381 tcttgctagg atacagttct cacatcacat ccgaacataa acaaccatgg gtaccactct
     1441 tgacgacacg gcttaccggt accgcaccag tgtcccgggg gacgccgagg ccatcgaggc
     1501 actggatggg tccttcacca ccgacaccgt cttccgcgtc accgccaccg gggacggctt
     1561 caccctgcgg gaggtgccgg tggacccgcc cctgaccaag gtgttccccg acgacgaatc
     1621 ggacgacgaa tcggacgacg gggaggacgg cgacccggac tcccggacgt tcgtcgcgta
     1681 cggggacgac ggcgacctgg cgggcttcgt ggtcgtctcg tactccggct ggaaccgccg
     1741 gctgaccgtc gaggacatcg aggtcgcccc ggagcaccgg gggcacgggg tcgggcgcgc
     1801 gttgatgggg ctcgcgacgg agttcgcccg cgagcggggc gccgggcacc tctggctgga
     1861 ggtcaccaac gtcaacgcac cggcgatcca cgcgtaccgg cggatggggt tcaccctctg
     1921 cggcctggac accgccctgt acgacggcac cgcctcggac ggcgagcagg cgctctacat
     1981 gagcatgccc tgcccctaat cagtactgac aataaaaaga ttcttgtttt caagaacttg
     2041 tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt
     2101 tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta
     2161 atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa
     2221 cgccgccatc cagtttaaac gagctcgaat tctaggcttc ctcgctcact gactcgctgc
     2281 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2341 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2401 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2461 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2521 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2581 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     2641 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2701 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2761 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2821 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     2881 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2941 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3001 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3061 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3121 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3181 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3241 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3301 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3361 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3421 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3481 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3541 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3601 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3661 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3721 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3781 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3841 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3901 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3961 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4021 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4081 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4141 aaataggggt tccgcgcaca tttccccgaa aagtgccacc t
//
