LOCUS       sequence-146371-        4031 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 4031)
  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 4031)
  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: pNX3s2-PK3
FEATURES             Location/Qualifiers
     source          1..4031
                     /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      564..751
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(619..643)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     polyA_signal    771..852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     796..815
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(866..1240)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVCVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(1259..1306)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(1403..1427)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1424..1627)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1453..1473)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1628..1931
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(2005..2024)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2006..2024)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2111..2128)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2282..2870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2362..2381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3041..3832,3833..3901))
                     /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     3664..3683
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3902..4006)
                     /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 tgaggcgcgc cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat
      601 aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa
      661 aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagtatg
      721 aggtcgctct tattgaccac acctctaccg gcagatccgc taggagatct gcagtgaaaa
      781 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
      841 aataaacaag tttcgaggtc gagtgtcagt cctgctcctc ggccacgaag tgcacgcagt
      901 tgccggccgg gtcgcgcagg gcgaactccc gcccccacgg ctgctcgccg atctcggtca
      961 tggccggccc ggaggcgtcc cggaagttcg tggacacgac ctccgaccac tcggcgtaca
     1021 gctcgtccag gccgcgcaca cacacccagg ccagggtgtt gtccggcacc acctggtcct
     1081 ggaccgcgct gatgaacagg gtcacgtcgt cccggaccac accggcgaag tcgtcctcca
     1141 cgaagtcccg ggagaacccg agccggtcgg tccagaactc gaccgctccg gcgacgtcgc
     1201 gcgcggtgag caccggaacg gcactggtca acttggccat ggtttagttc ctcaccttgt
     1261 cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgg tccgttccaa
     1321 tgcaccgttc ccggccgcgg aggctggatc ggtcccggtg tcttctatgg aggtcaaaac
     1381 agcgtggatg gcgtctccag gcgatctgac ggttcactaa acgagctctg cttatataga
     1441 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
     1501 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     1561 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     1621 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     1681 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     1741 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     1801 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     1861 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     1921 gttatgtaac ggtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct
     1981 atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg
     2041 ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2101 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2161 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2221 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2281 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2341 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2401 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2461 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2521 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2581 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2641 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2701 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     2761 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2821 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2881 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2941 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3001 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3061 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3121 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3181 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3241 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3301 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3361 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3421 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3481 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3541 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3601 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3661 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3721 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3781 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3841 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3901 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3961 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4021 aagtgccacc t
//
