LOCUS       sequence-62031-e        4678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized mKO2
            with SpHis5 selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4678)
  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 4678)
  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-SpHis5
FEATURES             Location/Qualifiers
     source          1..4678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      18..205
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(73..97)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            280..1480
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        280..623
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             624..1277
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      1283..1480
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1584..1603)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1585..1603)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1690..1707)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1861..2449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1941..1960)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2620..3411,3412..3480))
                     /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     3243..3262
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3481..3585)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3653..3671
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3709..3731)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3831..3850
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3931..3949
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3931..3948
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4019..4675
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
ORIGIN
        1 gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta
       61 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct
      121 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg
      181 ctcttattga ccacacctct accggcagat ccgctaggga taacagggta atatagatct
      241 gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc
      301 ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt
      361 tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg
      421 cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca
      481 gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg
      541 ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac
      601 atcacatccg aacataaaca accatgggta ggagggcttt tgtagaaaga aatacgaacg
      661 aaacgaaaat cagcgttgcc atcgctttgg acaaagctcc cttacctgaa gagtcgaatt
      721 ttattgatga acttataact tccaagcatg caaaccaaaa gggagaacaa gtaatccaag
      781 tagacacggg aattggattc ttggatcaca tgtatcatgc actggctaaa catgcaggct
      841 ggagcttacg actttactca agaggtgatt taatcatcga tgatcatcac actgcagaag
      901 atactgctat tgcacttggt attgcattca agcaggctat gggtaacttt gccggcgtta
      961 aaagatttgg acatgcttat tgtccacttg acgaagctct ttctagaagc gtagttgact
     1021 tgtcgggacg gccctatgct gttatcgatt tgggattaaa gcgtgaaaag gttggggaat
     1081 tgtcctgtga aatgatccct cacttactat attccttttc ggtagcagct ggaattactt
     1141 tgcatgttac ctgcttatat ggtagtaatg accatcatcg tgctgaaagc gcttttaaat
     1201 ctctggctgt tgccatgcgc gcggctacta gtcttactgg aagttctgaa gtcccaagca
     1261 cgaagggagt gttgtaaaga gtactgacaa taaaaagatt cttgttttca agaacttgtc
     1321 atttgtatag tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta
     1381 tatttttttt cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat
     1441 atcatgcgtc aatcgtatgt gaatgctggt cgctatactg ctgtcgattc gatactaacg
     1501 ccgccatcca gtttaaacga gctcgaattc atcgatgata tcagatccac tagtggccta
     1561 tgcggccgcg gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt
     1621 taacctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1681 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1741 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1801 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1861 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1921 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1981 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2041 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2101 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2161 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2221 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2281 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     2341 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2401 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2461 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2521 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2581 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2641 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2701 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2761 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2821 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2881 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     2941 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3001 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3061 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3121 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3181 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3241 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3301 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3361 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3421 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3481 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3541 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3601 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     3661 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     3721 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     3781 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     3841 gcagattgta ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta
     3901 atacataacc ttatgtatca tacacatacg atttaggtga cactatagaa cgcggccgcc
     3961 agctgaagct tcgtacgctg caggtcgacg gatcggtgac ggtgctggtt taattaacat
     4021 ggtgtccgta attaagccag aaatgaagat gaggtattac atggatggtt ctgtcaatgg
     4081 acacgagttt actattgaag gtgagggtac gggcagacct tacgaaggcc atcaagagat
     4141 gacattaaga gtaactatgg ctgaaggagg tccaatgcct ttcgctttcg acttagtttc
     4201 tcatgtgttt tgttacgggc atcgtgtgtt tactaagtac cctgaagaga taccagatta
     4261 ctttaaacaa gccttcccag agggactttc atgggaaaga tcccttgaat ttgaagatgg
     4321 agggagtgct tcagtttctg cacatatcag cttgagaggt aacacatttt accacaaatc
     4381 aaaattcaca ggcgtcaact tccctgccga tgggccaatc atgcaaaatc aatctgtcga
     4441 ctgggaacca tcaacagaga aaattacagc atctgatggt gttctcaaag gtgacgttac
     4501 catgtatttg aaactagaag gaggtggtaa tcacaaatgc caaatgaaaa caacttataa
     4561 ggccgctaag gaaatattgg aaatgccagg cgaccattac atcggtcata gactggttag
     4621 aaagaccgaa ggcaatatca ctgaacaggt tgaagatgca gtagcgcaca gttaaggc
//
