LOCUS       sequence-62149-e        4846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            mKeima with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4846)
  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 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: "PLoS One.";
            date: "2013-07-2"; volume: "8(7):e67902. doi"; pages: " 
            10.1371/journal.pone.0067902. Print 2013"
COMMENT     Sequence Label: pFA6a-link-yomKeima-Kan
FEATURES             Location/Qualifiers
     source          1..4846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..38
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(76..98)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     198..217
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        298..316
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     298..315
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      1075..1262
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1130..1154)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1337..2693
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1337..1680
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1681..2490
                     /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(1748..1767)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2496..2693
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2797..2816)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2798..2816)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2903..2920)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3074..3662)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3154..3173)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3833..4624,4625..4693))
                     /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     4456..4475
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4694..4798)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
       61 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
      121 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
      181 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat
      241 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
      301 taggtgacac tatagaacgc ggccgccagc tgaagcttcg tacgctgcag gtcgacggat
      361 cggtgacggt gctggtttaa ttaacatggt ttctgtgatc gctaaacaaa tgacttataa
      421 agtctacatg agtggaacag taaacggcca ttacttcgag gtagaaggtg atggtaaagg
      481 aaaaccatac gaaggcgaac aaaccgtgaa attgactgtt accaaaggag gaccactccc
      541 atttgcctgg gatatccttt cacctcaatt gcaatacggt agtataccat tcacaaaata
      601 tcctgaagat attccagact actttaaaca atctttccca gaaggatata catgggagag
      661 atccatgaat ttcgaagatg gtgctgtttg tacggtgtca aatgactctt ctatacaggg
      721 caattgtttt atctacaacg ttaagatttc aggcgagaat tttcctccaa atggtccagt
      781 aatgcaaaag aaaacacagg ggtgggaacc atctactgaa aggttatttg ccagagatgg
      841 tatgcttatt ggtaacgact acatggcact aaagctggaa gggggtggtc attacttatg
      901 tgagtttaag tcaacttata aggcgaaaaa gcctgtcaga atgcctggca gacacgagat
      961 cgaccgtaag ttggatgtta ctagccacaa cagagattac acatccgtcg aacaatgcga
     1021 aattgctatc gcaagacatt ctctattagg gtaaggcgcg ccacttctaa ataagcgaat
     1081 ttcttatgat ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa
     1141 ttttaaagtg actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg
     1201 taggtcaggt tgctttctca ggtatagtat gaggtcgctc ttattgacca cacctctacc
     1261 ggcagatccg ctagggataa cagggtaata tagatctgtt tagcttgcct cgtccccgcc
     1321 gggtcacccg gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg
     1381 cagctcaggg gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata
     1441 atcatttgca tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc
     1501 gctgcagacc tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc
     1561 cgcgcccctg tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat
     1621 acttcctttt aaaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc
     1681 atgggtaagg aaaagactca cgtttcgagg ccgcgattaa attccaacat ggatgctgat
     1741 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga
     1801 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     1861 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     1921 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     1981 ggcaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     2041 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     2101 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     2161 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     2221 cataagcttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     2281 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     2341 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     2401 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     2461 tttcatttga tgctcgatga gtttttctaa tcagtactga caataaaaag attcttgttt
     2521 tcaagaactt gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg
     2581 ttagcgtgat ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg
     2641 cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga
     2701 ttcgatacta acgccgccat ccagtttaaa cgagctcgaa ttcatcgatg atatcagatc
     2761 cactagtggc ctatgcggcc gcggatctgc cggtctccct atagtgagtc gtattaattt
     2821 cgataagcca ggttaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2881 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2941 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3001 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3061 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3121 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3181 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3241 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     3301 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3361 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3421 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3481 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     3541 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3601 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3661 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3721 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3781 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3841 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3901 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3961 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4021 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4081 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4141 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4201 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4261 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4321 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4381 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4441 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4501 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4561 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4621 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4681 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4741 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4801 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattat
//
