LOCUS       sequence-227916-        4735 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            mRuby2 with SpHis5 selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4735)
  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 4735)
  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-yomRuby2-SpHis5
FEATURES             Location/Qualifiers
     source          1..4735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             53..763
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     terminator      787..974
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(842..866)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1049..2249
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        1049..1392
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1393..2046
                     /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="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHTNQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMSNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      2052..2249
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2353..2372)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2354..2372)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2459..2476)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2630..3218)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2710..2729)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3389..4180,4181..4249))
                     /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     4012..4031
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4250..4354)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4422..4440
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4478..4500)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4600..4619
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4700..4718
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4700..4717
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 agcttcgtac gctgcaggtc gacggatcgg tgacggtgct ggtttaatta acatggtgtc
       61 caaaggagag gagttaatca aggaaaacat gagaatgaaa gttgtcatgg agggctccgt
      121 taatggtcac caattcaagt gtacagggga aggtgaaggt aatccttaca tgggtacaca
      181 aactatgaga attaaagtaa ttgaaggcgg accactacca tttgcatttg acattctggc
      241 aacgtcattc atgtacggat cacgaacttt catcaagtac cctaaaggta taccagactt
      301 tttcaagcaa tcttttccag agggttttac atgggaaagg gttacaagat acgaagatgg
      361 gggtgtcgtc acagttatgc aagatacttc attagaagat ggctgccttg tctatcatgt
      421 gcaagtaaga ggggtgaatt ttccttctaa cggacctgtg atgcagaaaa agaccaaagg
      481 ttgggaacca aatactgaaa tgatgtaccc agctgatgga ggtttgagag gctacacaca
      541 catggcgctt aaagttgatg gtggaggtca tttgtcttgt agttttgtta ccacttatcg
      601 ttctaaaaag actgttggca atatcaaaat gccaggaata catgctgtag accacagact
      661 agaaagactc gaagagagcg ataacgaaat gttcgttgta cagagagagc atgccgtagc
      721 caaatttgct ggcttaggcg gtggtatgga tgaattgtat aagtaaggcg cgccacttct
      781 aaataagcga atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata
      841 agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt
      901 aactctttcc tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac
      961 cacacctcta ccggcagatc cgctagggat aacagggtaa tatagatctg tttagcttgc
     1021 ctcgtccccg ccgggtcacc cggccagcga catggaggcc cagaataccc tccttgacag
     1081 tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca
     1141 tccccatgta taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa
     1201 ttacggctcc tcgctgcaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa
     1261 ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt
     1321 cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga
     1381 acataaacaa ccatgggtag gagggctttt gtagaaagaa atacgaacga aacgaaaatc
     1441 agcgttgcca tcgctttgga caaagctccc ttacctgaag agtcgaattt tattgatgaa
     1501 cttataactt ccaagcatac aaaccaaaag ggagaacaag taatccaagt agacacggga
     1561 attggattct tggatcacat gtatcatgca ctggctaaac atgcaggctg gagcttacga
     1621 ctttactcaa gaggtgattt aatcatcgat gatcatcaca ctgcagaaga tactgctatt
     1681 gcacttggta ttgcattcaa gcaggctatg agtaactttg ccggcgttaa aagatttgga
     1741 catgcttatt gtccacttga cgaagctctt tctagaagcg tagttgactt gtcgggacgg
     1801 ccctatgctg ttatcgattt gggattaaag cgtgaaaagg ttggggaatt gtcctgtgaa
     1861 atgatccctc acttactata ttccttttcg gtagcagctg gaattacttt gcatgttacc
     1921 tgcttatatg gtagtaatga ccatcatcgt gctgaaagcg cttttaaatc tctggctgtt
     1981 gccatgcgcg cggctactag tcttactgga agttctgaag tcccaagcac gaagggagtg
     2041 ttgtaaagag tactgacaat aaaaagattc ttgttttcaa gaacttgtca tttgtatagt
     2101 ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat attttttttc
     2161 gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca
     2221 atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc cgccatccag
     2281 tttaaacgag ctcgaattca tcgatgatat cagatccact agtggcctat gcggccgcgg
     2341 atctgccggt ctccctatag tgagtcgtat taatttcgat aagccaggtt aacctgcatt
     2401 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2461 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2521 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2581 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2641 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2701 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2761 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2821 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2881 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2941 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3001 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3061 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3121 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3181 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3241 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3301 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3361 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3421 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3481 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3541 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3601 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3661 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3721 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3781 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3841 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3901 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3961 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4021 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4081 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4141 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4201 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4261 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4321 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4381 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     4441 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     4501 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     4561 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     4621 tgagagtgca ccatatggac atattgtcgt tagaacgcgg ctacaattaa tacataacct
     4681 tatgtatcat acacatacga tttaggtgac actatagaac gcggccgcca gctga
//
