LOCUS       sequence-149819-        4531 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 4531)
  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 4531)
  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: pNX3c2-Myc13
FEATURES             Location/Qualifiers
     source          1..4531
                     /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             418..915
                     /codon_start=1
                     /product="13 tandem Myc epitope tags"
                     /label=13xMyc
                     /translation="EQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLIS
                     EEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLD
                     GEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISE
                     EDL"
     terminator      978..1165
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1033..1057)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     enhancer        1190..1493
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1494..1697
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1648..1668
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1742..1761
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1742..1760
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1776..2348
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      2354..2551
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2611..2628)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2782..3370)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2862..2881)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3541..4332,4333..4401))
                     /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     4164..4183
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4402..4506)
                     /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 ctagcgcgat taacggtgaa
      421 caaaagctaa tctccgagga agacttgaac ggtgaacaaa aattaatctc agaagaagac
      481 ttgaacggac tcgacggtga acaaaagttg atttctgaag aagatttgaa cggtgaacaa
      541 aagctaatct ccgaggaaga cttgaacggt gaacaaaaat taatctcaga agaagacttg
      601 aacggactcg acggtgaaca aaagttgatt tctgaagaag atttgaacgg tgaacaaaag
      661 ctaatctccg aggaagactt gaacggtgaa caaaaattaa tctcagaaga agacttgaac
      721 ggactcgacg gtgaacaaaa gttgatttct gaagaagatt tgaacggtga acaaaagcta
      781 atctccgagg aagacttgaa cggtgaacaa aaattaatct cagaagaaga cttgaacgga
      841 ctcgacggtg aacaaaagtt gatttctgaa gaagatttga acggtgaaca aaagctaatc
      901 tccgaggaag acttgaacgg tgaacaaaaa ttaatcaatc agtacaagtc tagatgaggc
      961 gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta
     1021 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct
     1081 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg
     1141 ctcttattga ccacacctct accggcagat ccgctaggag atctcgcgac gttacataac
     1201 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     1261 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     1321 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     1381 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     1441 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     1501 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     1561 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     1621 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     1681 tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa
     1741 ttaatacgac tcactatagg gagacccaag cttacatggg taccactctt gacgacacgg
     1801 cttaccggta ccgcaccagt gtcccggggg acgccgaggc catcgaggca ctggatgggt
     1861 ccttcaccac cgacaccgtc ttccgcgtca ccgccaccgg ggacggcttc accctgcggg
     1921 aggtgccggt ggacccgccc ctgaccaagg tgttccccga cgacgaatcg gacgacgaat
     1981 cggacgacgg ggaggacggc gacccggact cccggacgtt cgtcgcgtac ggggacgacg
     2041 gcgacctggc gggcttcgtg gtcgtctcgt actccggctg gaaccgccgg ctgaccgtcg
     2101 aggacatcga ggtcgccccg gagcaccggg ggcacggggt cgggcgcgcg ttgatggggc
     2161 tcgcgacgga gttcgcccgc gagcggggcg ccgggcacct ctggctggag gtcaccaacg
     2221 tcaacgcacc ggcgatccac gcgtaccggc ggatggggtt caccctctgc ggcctggaca
     2281 ccgccctgta cgacggcacc gcctcggacg gcgagcaggc gctctacatg agcatgccct
     2341 gcccctaatc agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata
     2401 gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt
     2461 tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt
     2521 caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc
     2581 agtgttaaac gagctcgaat tctaggcttc ctcgctcact gactcgctgc gctcggtcgt
     2641 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2701 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2761 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2821 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2881 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2941 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     3001 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3061 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3121 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3181 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     3241 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3301 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3361 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3421 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3481 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3541 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3601 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3661 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3721 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3781 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3841 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3901 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3961 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4021 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4081 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4141 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4201 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4261 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4321 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4381 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4441 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4501 tccgcgcaca tttccccgaa aagtgccacc t
//
