LOCUS       sequence-172882-        4846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tag S. pombe genes in C terminal with mEOS3.2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4846)
  AUTHORS   Laplante C, Huang F, Tebbs IR, Bewersdorf J, Pollard TD
  TITLE     Molecular organization of cytokinesis nodes and contractile rings by
            super-resolution fluorescence microscopy of live fission yeast.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Oct 4;113(40):E5876-E5885. Epub 2016 
            Sep 19.
  PUBMED    27647921
REFERENCE   2  (bases 1 to 4846)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2016-10-4"; volume: "113(40)"; pages: 
            "E5876-E5885. Epub 2016 Sep 19"
COMMENT     Sequence Label: pFA6a-mEOS3.2-KanMX6
FEATURES             Location/Qualifiers
     source          1..4846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             63..740
                     /codon_start=1
                     /product="improved monomeric variant of green-to-red 
                     photoswitchable fluorescent protein EosFP (Zhang et al., 
                     2012)"
                     /label=mEos3.2
                     /translation="SAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVKE
                     GGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNARN
                     DITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALLLE
                     GNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPDN
                     ARR"
     terminator      761..948
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(816..840)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1023..2379
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1023..1366
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1367..2176
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1434..1453)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2182..2379
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2483..2502)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2484..2502)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2589..2606)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2760..3348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2840..2859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3519..4310,4311..4379))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGSRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4142..4161
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4380..4484)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4552..4570
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4608..4630)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4730..4749
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4830..4846,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4830..4846,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acagtgcgat taagccagac atgaagatca aactccgtat ggaaggcaac gtaaacgggc
      121 accactttgt gatcgacgga gatggtacag gcaagccttt tgagggaaaa cagagtatgg
      181 atcttgaagt caaagagggc ggacctctgc cttttgcctt tgatatcctg accactgcat
      241 tccattacgg caacagggta ttcgccaaat atccagacaa catacaagac tattttaagc
      301 agtcgtttcc taaggggtat tcgtgggaac gaagcttgac tttcgaagac gggggcattt
      361 gcaacgccag aaacgacata acaatggaag gggacacttt ctataataaa gttcgatttt
      421 atggtaccaa ctttcccgcc aatggtccag ttatgcagaa gaagacgctg aaatgggagc
      481 cctccactga gaaaatgtat gtgcgtgatg gagtgctgac gggtgatatt gagatggctt
      541 tgttgcttga aggaaatgcc cattaccgat gtgacttcag aactacttac aaagctaagg
      601 agaagggtgt caagttacca ggcgcccact ttgtggacca ctgcattgag attttaagcc
      661 atgacaaaga ttacaacaag gttaagctgt atgagcatgc tgttgctcat tctggattgc
      721 ctgacaatgc cagacgataa ggcgcgccac ttctaaataa gcgaatttct tatgatttat
      781 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
      841 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
      901 ttctcaggta tagtattagg tcgctcttat tgaccacacc tctaccggca gatccgctag
      961 ggataacagg gtaatataga tctgtttagc ttgcctcgtc cccgccgggt cacccggcca
     1021 gcgacatgga ggcccagaat accctccttg acagtcttga cgtgcgcagc tcaggggcat
     1081 gatgtgactg tcgcccgtac atttagccca tacatcccca tgtataatca tttgcatcca
     1141 tacattttga tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg cagacctgct
     1201 agcagggaaa cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga
     1261 gaaatataaa aggttaggat ttgccactga ggttcttctt tcatatactt ccttttaaaa
     1321 tcttgctagg atacagttct cacatcacat ccgaacataa acaaccatgg gtaaggaaaa
     1381 gactcacgtt tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     1441 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc
     1501 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     1561 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     1621 tatccgtact cctgatgatg catggttact caccactgcg atccccggca aaacagcatt
     1681 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     1741 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     1801 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     1861 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata agcttttgcc
     1921 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     1981 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2041 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     2101 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     2161 cgatgagttt ttctaatcag tactgacaat aaaaagattc ttgttttcaa gaacttgtca
     2221 tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat
     2281 attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata
     2341 tcatgcgtca atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc
     2401 cgccatccag tttaaacgag ctcgaattca tcgatgatat cagatccact agtggcctat
     2461 gcggccgcgg atctgccggt ctccctatag tgagtcgtat taatttcgat aagccaggtt
     2521 aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2581 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2641 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2701 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2761 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2821 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2881 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2941 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3001 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3061 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3121 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3181 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3241 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3301 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3361 atcttttcta cggggtctta cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3421 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3481 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3541 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3601 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3661 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3721 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3781 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3841 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3901 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3961 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4021 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4081 aagtcattct gagaatagtg tatgcggcta ccgagttgct cttgcccggc gtcaatacgg
     4141 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4201 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4261 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4321 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4381 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4441 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4501 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     4561 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     4621 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     4681 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     4741 cagattgtac tgagagtgca ccatatggac atattgtcgt tagaacgcgg ctacaattaa
     4801 tacataacct tatgtatcat acacatacga tttaggtgac actata
//
