LOCUS       sequence-146331-        4906 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 4906)
  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 4906)
  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: pNX3a2-Myc13
FEATURES             Location/Qualifiers
     source          1..4906
                     /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"
     polyA_signal    1185..1266
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1210..1229
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1285..2079)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1396..1415)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2006..2025
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2134..2181)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(2278..2302)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2299..2502)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2328..2348)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2503..2806
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(2880..2899)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2881..2899)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2986..3003)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3157..3745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3237..3256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3916..4707,4708..4776))
                     /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     4539..4558
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4777..4881)
                     /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 atctgcagtg aaaaaaatgc
     1201 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     1261 caagttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
     1321 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
     1381 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
     1441 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
     1501 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
     1561 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
     1621 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
     1681 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     1741 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     1801 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     1861 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
     1921 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     1981 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     2041 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc cagctttttg
     2101 caaaagccta gccatggttt agttcctcac cttgtcgtat tatactatgc cgatatacta
     2161 tgccgatgat taattgtcaa cacggtccgt tccaatgcac cgttcccggc cgcggaggct
     2221 ggatcggtcc cggtgtcttc tatggaggtc aaaacagcgt ggatggcgtc tccaggcgat
     2281 ctgacggttc actaaacgag ctctgcttat atagacctcc caccgtacac gcctaccgcc
     2341 catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg
     2401 ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa
     2461 ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac
     2521 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     2581 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
     2641 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     2701 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     2761 gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacggttt aaacgagctc
     2821 gaattcatcg atgatatcag atccactagt ggcctatgcg gccgcggatc tgccggtctc
     2881 cctatagtga gtcgtattaa tttcgataag ccaggttaac ctgcattaat gaatcggcca
     2941 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3001 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3061 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3121 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3181 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3241 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3301 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     3361 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3421 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3481 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3541 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     3601 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3661 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3721 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3781 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3841 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3901 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3961 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4021 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4081 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4141 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4201 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4261 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4321 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4381 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4441 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4501 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4561 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4621 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4681 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4741 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4801 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4861 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacct
//
