LOCUS       sequence-62234-e        4861 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            mRuby with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4861)
  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 4861)
  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-yomRuby-Kan
FEATURES             Location/Qualifiers
     source          1..4861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     54..73
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        154..172
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     154..171
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      946..1133
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1001..1025)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1208..2564
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1208..1551
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1552..2361
                     /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(1619..1638)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2367..2564
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2668..2687)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2669..2687)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2774..2791)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2945..3533)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3025..3044)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3704..4495,4496..4564))
                     /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     4327..4346
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4565..4669)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4737..4755
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4793..4815)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
       61 agagcagatt gtactgagag tgcaccatat ggacatattg tcgttagaac gcggctacaa
      121 ttaatacata accttatgta tcatacacat acgatttagg tgacactata gaacgcggcc
      181 gccagctgaa gcttcgtacg ctgcaggtcg acggatcggt gacggtgctg gtttaattaa
      241 catgaactcc ttgattaagg aaaatatgag aatgaaagtt gtgttagaag gttccgtaaa
      301 cggtcaccag tttaagtgta caggcgaagg agaaggaaac ccttacatgg ggacacaaac
      361 tatgagaatc aaggtaatcg aaggtggtcc tcttccattt gccttcgaca tattggcgac
      421 atctttcatg tacggctctc gaactttcat caaataccca aaagggattc ctgacttttt
      481 caaacaatca tttccagaag gcttcacctg ggaaagggtt acaagatacg aggatggtgg
      541 cgttattact gtaatgcaag atacttcact ggaggatggt tgtctcgtct accacgctca
      601 agtaagaggt gtcaattttc cttctaatgg agctgtgatg cagaaaaaga ccaaaggttg
      661 ggaaccaaat acagagatga tgtatccagc cgatggggga ctaagaggat atacgcacat
      721 ggctcttaag gtcgatggcg gaggtcatct aagctgctca tttgttacaa cttacagaag
      781 caaaaagact gttggcaaca tcaaaatgcc aggtatacat gcagttgacc atcgtttgga
      841 aagattagag gaaagtgata atgagatgtt tgttgtgcaa agagaacatg cagtcgcaaa
      901 atttgctggc ttaggtggtg ggtaaggcgc gccacttcta aataagcgaa tttcttatga
      961 tttatgattt ttattattaa ataagttata aaaaaaataa gtgtatacaa attttaaagt
     1021 gactcttagg ttttaaaacg aaaattctta ttcttgagta actctttcct gtaggtcagg
     1081 ttgctttctc aggtatagta tgaggtcgct cttattgacc acacctctac cggcagatcc
     1141 gctagggata acagggtaat atagatctgt ttagcttgcc tcgtccccgc cgggtcaccc
     1201 ggccagcgac atggaggccc agaataccct ccttgacagt cttgacgtgc gcagctcagg
     1261 ggcatgatgt gactgtcgcc cgtacattta gcccatacat ccccatgtat aatcatttgc
     1321 atccatacat tttgatggcc gcacggcgcg aagcaaaaat tacggctcct cgctgcagac
     1381 ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct
     1441 gtagagaaat ataaaaggtt aggatttgcc actgaggttc ttctttcata tacttccttt
     1501 taaaatcttg ctaggataca gttctcacat cacatccgaa cataaacaac catgggtaag
     1561 gaaaagactc acgtttcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     1621 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg
     1681 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     1741 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     1801 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggcaaaaca
     1861 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     1921 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     1981 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     2041 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataagctt
     2101 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     2161 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     2221 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     2281 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     2341 atgctcgatg agtttttcta atcagtactg acaataaaaa gattcttgtt ttcaagaact
     2401 tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga
     2461 tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag
     2521 taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact
     2581 aacgccgcca tccagtttaa acgagctcga attcatcgat gatatcagat ccactagtgg
     2641 cctatgcggc cgcggatctg ccggtctccc tatagtgagt cgtattaatt tcgataagcc
     2701 aggttaacct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2761 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2821 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2881 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2941 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3001 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3061 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3121 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     3181 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3241 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3301 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3361 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     3421 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3481 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3541 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3601 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3661 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3721 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3781 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3841 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3901 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3961 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4021 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4081 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4141 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4201 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4261 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4321 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4381 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4441 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4501 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4561 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4621 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4681 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     4741 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     4801 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     4861 c
//
