LOCUS       sequence-62151-e        4912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            TagRFP-T with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4912)
  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 4912)
  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-yoTagRFP-T-Kan
FEATURES             Location/Qualifiers
     source          1..4912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..134
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        215..233
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     215..232
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      1058..1245
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1113..1137)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1320..2676
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1320..1663
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1664..2473
                     /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(1731..1750)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2479..2676
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2780..2799)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2781..2799)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2886..2903)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3057..3645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3137..3156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3816..4607,4608..4676))
                     /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     4439..4458
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4677..4781)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4849..4867
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(4905..4912,1..15))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
       61 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
      121 cagagcagat tgtactgaga gtgcaccata tggacatatt gtcgttagaa cgcggctaca
      181 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaacgcggc
      241 cgccagctga agcttcgtac gctgcaggtc gacggatcgg tgacggtgct ggtttaatta
      301 acatggtatc taaaggtgaa gagttgataa aggaaaatat gcacatgaaa ctctatatgg
      361 aaggcacagt gaataaccat cattttaagt gtacttcaga aggagaaggt aagccttatg
      421 aggggacaca aactatgaga attaaagttg ttgaaggagg tccactacca ttcgcttttg
      481 acatcctggc aacgagcttc atgtatggct ccagaacatt cataaaccat acacaaggta
      541 ttccagattt cttcaaacaa agttttccag aaggctttac ttgggagaga gttactacct
      601 acgaggacgg gggtgtttta actgcgacac aggatacatc attacaagat ggctgtttga
      661 tctacaacgt aaaaatcaga ggtgtcaatt ttccatctaa cggacctgtt atgcagaaaa
      721 agaccttggg ttgggaggct aatactgaaa tgctataccc agcagatgga ggattggaag
      781 gtaggacaga tatggcattg aaacttgtag gaggcggtca cctaatttgc aactttaaga
      841 caacctacag aagcaaaaag cctgctaaaa atcttaaaat gccaggggtg tactacgtgg
      901 atcatagact tgaacgtatc aaggaagctg acaaagagac ttacgttgaa caacacgaag
      961 tcgccgtcgc cagatactgt gacttacctt caaagttagg tcataaactg aatggtatgg
     1021 atgaattgta taagtaaggc gcgccacttc taaataagcg aatttcttat gatttatgat
     1081 ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta
     1141 ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc
     1201 tcaggtatag tatgaggtcg ctcttattga ccacacctct accggcagat ccgctaggga
     1261 taacagggta atatagatct gtttagcttg cctcgtcccc gccgggtcac ccggccagcg
     1321 acatggaggc ccagaatacc ctccttgaca gtcttgacgt gcgcagctca ggggcatgat
     1381 gtgactgtcg cccgtacatt tagcccatac atccccatgt ataatcattt gcatccatac
     1441 attttgatgg ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag acctgcgagc
     1501 agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc ctgtagagaa
     1561 atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct tttaaaatct
     1621 tgctaggata cagttctcac atcacatccg aacataaaca accatgggta aggaaaagac
     1681 tcacgtttcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     1741 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga
     1801 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     1861 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     1921 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggcaaaa cagcattcca
     1981 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2041 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     2101 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     2161 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt
     2221 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     2281 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     2341 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     2401 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     2461 tgagtttttc taatcagtac tgacaataaa aagattcttg ttttcaagaa cttgtcattt
     2521 gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt
     2581 ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca
     2641 tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata ctaacgccgc
     2701 catccagttt aaacgagctc gaattcatcg atgatatcag atccactagt ggcctatgcg
     2761 gccgcggatc tgccggtctc cctatagtga gtcgtattaa tttcgataag ccaggttaac
     2821 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2881 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2941 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3001 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3061 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3121 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3181 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3241 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3301 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3361 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3421 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3481 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3541 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3601 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3661 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3721 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3781 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3841 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3901 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3961 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4021 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4081 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4141 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4201 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4261 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4321 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4381 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4441 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4501 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4561 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4621 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4681 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4741 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4801 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4861 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg ac
//
