LOCUS       sequence-146328-        4459 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 4459)
  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 4459)
  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-HA3
FEATURES             Location/Qualifiers
     source          1..4459
                     /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             409..435
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             439..465
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             472..498
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      531..718
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(586..610)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     polyA_signal    738..819
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     763..782
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(838..1632)
                     /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(949..968)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1559..1578
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1687..1734)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(1831..1855)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1852..2055)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1881..1901)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2056..2359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(2433..2452)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2434..2452)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2539..2556)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2710..3298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2790..2809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3469..4260,4261..4329))
                     /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     4092..4111
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4330..4434)
                     /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 ctagcatgta cccatacgat
      421 gttcctgact atgcgggcta cccgtatgac gtgccggact atgcaggagc ctatccttat
      481 gacgttccag attacgctgg ttctagatga ggcgcgccac ttctaaataa gcgaatttct
      541 tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt
      601 aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg
      661 tcaggttgct ttctcaggta tagtatgagg tcgctcttat tgaccacacc tctaccggca
      721 gatccgctag gagatctgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
      781 gctttatttg taaccattat aagctgcaat aaacaagttc gaaccccaga gtcccgctca
      841 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
      901 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
      961 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
     1021 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac
     1081 gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag
     1141 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
     1201 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
     1261 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
     1321 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
     1381 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
     1441 tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
     1501 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
     1561 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
     1621 ttgttcaatc atgcgaaacg atccagcttt ttgcaaaagc ctagccatgg tttagttcct
     1681 caccttgtcg tattatacta tgccgatata ctatgccgat gattaattgt caacacggtc
     1741 cgttccaatg caccgttccc ggccgcggag gctggatcgg tcccggtgtc ttctatggag
     1801 gtcaaaacag cgtggatggc gtctccaggc gatctgacgg ttcactaaac gagctctgct
     1861 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt
     1921 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     1981 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     2041 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     2101 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     2161 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     2221 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     2281 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     2341 taccgtaagt tatgtaacgg tttaaacgag ctcgaattca tcgatgatat cagatccact
     2401 agtggcctat gcggccgcgg atctgccggt ctccctatag tgagtcgtat taatttcgat
     2461 aagccaggtt aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2521 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2581 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2641 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2701 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2761 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2821 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2881 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     2941 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3001 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3061 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3121 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     3181 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3241 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3301 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3361 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3421 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3481 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3541 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3601 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3661 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3721 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3781 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3841 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3901 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3961 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4021 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4081 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4141 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4201 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4261 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4321 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4381 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4441 tccccgaaaa gtgccacct
//
