LOCUS       sequence-62233-e        4879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            TagRFp657 with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4879)
  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 4879)
  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-yoTagRFP657-Kan
FEATURES             Location/Qualifiers
     source          1..4879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     125..143
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(181..203)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     303..322
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        403..421
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     403..420
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      1213..1400
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1268..1292)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1475..2831
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1475..1818
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1819..2628
                     /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(1886..1905)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2634..2831
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2935..2954)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2936..2954)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3041..3058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3212..3800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3292..3311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3971..4762,4763..4831))
                     /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     4594..4613
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4832..4879,1..57))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
       61 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
      121 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
      181 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
      241 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
      301 tgcggcatca gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg
      361 cggctacaat taatacataa ccttatgtat catacacata cgatttaggt gacactatag
      421 aacgcggccg ccagctgaag cttcgtacgc tgcaggtcga cggatcggtg acggtgctgg
      481 tttaattaac atgagcgaat tgatcacaga aaatatgcac atgaagttgt atatggaggg
      541 aaccgtcaat aaccatcatt tcaaatgcac ttctgaaggt gagggcaaac cttacgaagg
      601 gacccaaaca cagagaatta aggtagtaga agggggtcca ctgccatttg cctttgacat
      661 cctagcaaca tccttcatgt acggttccca cacattcata aaccacactc aaggaatacc
      721 agacttttgg aagcagagtt ttccagaagg ttttacgtgg gaaagagtta ctacttacga
      781 ggacggtggt gtattaacag ccacacaaga tacatcatta caagatggtt gtctgatcta
      841 caacgttaaa atcagaggtg ttaactttcc ttcaaatggg ccagttatgc aaaagaaaac
      901 tttaggctgg gaggcccata ctgaaatgct ttacccagca gatggcggct tggaaggcag
      961 gacggctttg gctttaaagc tagttggtgg aggacatttg atttgtaatt tcaaaacaac
     1021 ttacagaagc aaaaagcctg ctaaaaatct taagatgcca ggagtgtact atgtcgatta
     1081 cagattggaa cgtattaaag aggctgataa ggaaacctat gtggaacaac atgaagtggc
     1141 agtcgcgaga tattgtgatc ttccttctaa actaggtcat aaactcaatt aaggcgcgcc
     1201 acttctaaat aagcgaattt cttatgattt atgattttta ttattaaata agttataaaa
     1261 aaaataagtg tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc
     1321 ttgagtaact ctttcctgta ggtcaggttg ctttctcagg tatagtatga ggtcgctctt
     1381 attgaccaca cctctaccgg cagatccgct agggataaca gggtaatata gatctgttta
     1441 gcttgcctcg tccccgccgg gtcacccggc cagcgacatg gaggcccaga ataccctcct
     1501 tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt acatttagcc
     1561 catacatccc catgtataat catttgcatc catacatttt gatggccgca cggcgcgaag
     1621 caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc tcacagacgc
     1681 gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg atttgccact
     1741 gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt ctcacatcac
     1801 atccgaacat aaacaaccat gggtaaggaa aagactcacg tttcgaggcc gcgattaaat
     1861 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     1921 ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     1981 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     2041 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     2101 ctcaccactg cgatccccgg caaaacagca ttccaggtat tagaagaata tcctgattca
     2161 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     2221 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     2281 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     2341 caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt cgtcactcat
     2401 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     2461 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     2521 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     2581 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agtactgaca
     2641 ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg
     2701 ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc
     2761 cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg
     2821 tcgctatact gctgtcgatt cgatactaac gccgccatcc agtttaaacg agctcgaatt
     2881 catcgatgat atcagatcca ctagtggcct atgcggccgc ggatctgccg gtctccctat
     2941 agtgagtcgt attaatttcg ataagccagg ttaacctgca ttaatgaatc ggccaacgcg
     3001 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3061 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3121 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3181 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3241 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3301 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3361 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     3421 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3481 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3541 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3601 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     3661 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3721 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3781 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3841 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3901 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3961 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4021 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4081 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4141 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4201 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4261 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4321 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4381 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4441 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4501 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4561 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4621 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4681 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4741 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4801 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4861 tttatcaggg ttattgtct
//
