LOCUS       sequence-62239-e        4834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized mKO2
            with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4834)
  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 4834)
  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-yomKO2-Kan
FEATURES             Location/Qualifiers
     source          1..4834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     67..86
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        167..185
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     167..184
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             255..911
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     terminator      932..1119
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(987..1011)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1194..2550
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1194..1537
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1538..2347
                     /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(1605..1624)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2353..2550
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2654..2673)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2655..2673)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2760..2777)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2931..3519)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3011..3030)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3690..4481,4482..4550))
                     /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     4313..4332
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4551..4655)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4723..4741
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4779..4801)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
       61 actatgcggc atcagagcag attgtactga gagtgcacca tatggacata ttgtcgttag
      121 aacgcggcta caattaatac ataaccttat gtatcataca catacgattt aggtgacact
      181 atagaacgcg gccgccagct gaagcttcgt acgctgcagg tcgacggatc ggtgacggtg
      241 ctggtttaat taacatggtg tccgtaatta agccagaaat gaagatgagg tattacatgg
      301 atggttctgt caatggacac gagtttacta ttgaaggtga gggtacgggc agaccttacg
      361 aaggccatca agagatgaca ttaagagtaa ctatggctga aggaggtcca atgcctttcg
      421 ctttcgactt agtttctcat gtgttttgtt acgggcatcg tgtgtttact aagtaccctg
      481 aagagatacc agattacttt aaacaagcct tcccagaggg actttcatgg gaaagatccc
      541 ttgaatttga agatggaggg agtgcttcag tttctgcaca tatcagcttg agaggtaaca
      601 cattttacca caaatcaaaa ttcacaggcg tcaacttccc tgccgatggg ccaatcatgc
      661 aaaatcaatc tgtcgactgg gaaccatcaa cagagaaaat tacagcatct gatggtgttc
      721 tcaaaggtga cgttaccatg tatttgaaac tagaaggagg tggtaatcac aaatgccaaa
      781 tgaaaacaac ttataaggcc gctaaggaaa tattggaaat gccaggcgac cattacatcg
      841 gtcatagact ggttagaaag accgaaggca atatcactga acaggttgaa gatgcagtag
      901 cgcacagtta aggcgcgcca cttctaaata agcgaatttc ttatgattta tgatttttat
      961 tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt
     1021 aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt
     1081 atagtatgag gtcgctctta ttgaccacac ctctaccggc agatccgcta gggataacag
     1141 ggtaatatag atctgtttag cttgcctcgt ccccgccggg tcacccggcc agcgacatgg
     1201 aggcccagaa taccctcctt gacagtcttg acgtgcgcag ctcaggggca tgatgtgact
     1261 gtcgcccgta catttagccc atacatcccc atgtataatc atttgcatcc atacattttg
     1321 atggccgcac ggcgcgaagc aaaaattacg gctcctcgct gcagacctgc gagcagggaa
     1381 acgctcccct cacagacgcg ttgaattgtc cccacgccgc gcccctgtag agaaatataa
     1441 aaggttagga tttgccactg aggttcttct ttcatatact tccttttaaa atcttgctag
     1501 gatacagttc tcacatcaca tccgaacata aacaaccatg ggtaaggaaa agactcacgt
     1561 ttcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1621 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
     1681 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     1741 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     1801 tcctgatgat gcatggttac tcaccactgc gatccccggc aaaacagcat tccaggtatt
     1861 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     1921 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     1981 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2041 taatggctgg cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc
     2101 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2161 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2221 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2281 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     2341 tttctaatca gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag
     2401 tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt
     2461 cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc
     2521 aatcgtatgt gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca
     2581 gtttaaacga gctcgaattc atcgatgata tcagatccac tagtggccta tgcggccgcg
     2641 gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat
     2701 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2761 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2821 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2881 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2941 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3001 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3061 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3121 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3181 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3241 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3301 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3361 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3421 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3481 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3541 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3601 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3661 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3721 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3781 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3841 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3901 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3961 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4021 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4081 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4141 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4201 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4261 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4321 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4381 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4441 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4501 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4561 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4621 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4681 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     4741 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     4801 gagacggtca cagcttgtct gtaagcggat gccg
//
