LOCUS       sequence-62214-e        4858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            mEos2 with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4858)
  AUTHORS   Lee S, Lim WA, Thorn KS
  TITLE     Improved Blue, Green, and Red Fluorescent Protein Tagging Vectors 
            for S. cerevisiae.
  JOURNAL   PLoS One. 2013 Jul 2;8(7):e67902. doi: 10.1371/journal.pone.0067902.
            Print 2013.
  PUBMED    23844123
REFERENCE   2  (bases 1 to 4858)
  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: "PLoS One.";
            date: "2013-07-2"; volume: "8(7):e67902. doi"; pages: " 
            10.1371/journal.pone.0067902. Print 2013"
COMMENT     Sequence Label: pFA6a-link-yomEos2-Kan
FEATURES             Location/Qualifiers
     source          1..4858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..60)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     160..179
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        260..278
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     260..277
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      1049..1236
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1104..1128)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1311..2667
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1311..1654
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1655..2464
                     /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(1722..1741)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2470..2667
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2771..2790)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2772..2790)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2877..2894)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3048..3636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3128..3147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3807..4598,4599..4667))
                     /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     4430..4449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4668..4772)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4840..4858
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
       61 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
      121 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
      181 tgagagtgca ccatatggac atattgtcgt tagaacgcgg ctacaattaa tacataacct
      241 tatgtatcat acacatacga tttaggtgac actatagaac gcggccgcca gctgaagctt
      301 cgtacgctgc aggtcgacgg atcggtgacg gtgctggttt aattaacatg tctgccatta
      361 aacctgacat gaaaatcaaa ctgagaatgg aaggaaacgt caatggccat cactttgtaa
      421 tcgacggtga tggtacagga aaaccattcg aaggtaagca aagcatggat ctggaggtaa
      481 aagagggtgg cccactccca tttgcattcg atatcttaac caccgccttt cattacggta
      541 acagagtttt tgcaaagtat cctgataaca ttcaagatta tttcaaacag tcatttccaa
      601 agggttacag ttgggaaaga agcctaacat ttgaagatgg cggtatttgt atcgctagaa
      661 atgatattac aatggagggt gacactttct acaataaggt aagattttat ggaacaaatt
      721 tccctgctaa cggacctgtt atgcaaaaga aaacgttgaa gtgggaacca tctacagaga
      781 aaatgtacgt gcgtgacggg gttttaactg gtgatatcca catggcattg ctattagaag
      841 gcaatgcgca ctacagatgc gatttcagaa ctacttacaa agctaaggaa aagggggtga
      901 aacttccagg ataccacttt gttgaccatt gtatagaaat attgtcacat gataaggact
      961 acaacaaagt caaattgtat gaacatgctg ttgcccattc cggccttcca gataatgcta
     1021 ggcgataagg cgcgccactt ctaaataagc gaatttctta tgatttatga tttttattat
     1081 taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa
     1141 acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata
     1201 gtatgaggtc gctcttattg accacacctc taccggcaga tccgctaggg ataacagggt
     1261 aatatagatc tgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg
     1321 cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc
     1381 gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata cattttgatg
     1441 gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg
     1501 ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag
     1561 gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat
     1621 acagttctca catcacatcc gaacataaac aaccatgggt aaggaaaaga ctcacgtttc
     1681 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     1741 taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga
     1801 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     1861 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     1921 tgatgatgca tggttactca ccactgcgat ccccggcaaa acagcattcc aggtattaga
     1981 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     2041 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     2101 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     2161 tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga
     2221 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     2281 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     2341 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     2401 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     2461 ctaatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt tgtatagttt
     2521 ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat tttttttcgc
     2581 ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat
     2641 cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg ccatccagtt
     2701 taaacgagct cgaattcatc gatgatatca gatccactag tggcctatgc ggccgcggat
     2761 ctgccggtct ccctatagtg agtcgtatta atttcgataa gccaggttaa cctgcattaa
     2821 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2881 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2941 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3001 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3061 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3121 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3181 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3241 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3301 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3361 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3421 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3481 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3541 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3601 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3661 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3721 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3781 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3841 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3901 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3961 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4021 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4081 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4141 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4201 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4261 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4321 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4381 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4441 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4501 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4561 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4621 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4681 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4741 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4801 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggc
//
