LOCUS       sequence-62226-e        4891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            mRuby2 with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4891)
  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 4891)
  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-Kan
FEATURES             Location/Qualifiers
     source          1..4891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     104..122
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(160..182)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     282..301
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        382..400
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     382..399
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             470..1180
                     /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      1204..1391
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1259..1283)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1466..2822
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1466..1809
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1810..2619
                     /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(1877..1896)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2625..2822
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2926..2945)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2927..2945)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3032..3049)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3203..3791)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3283..3302)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3962..4753,4754..4822))
                     /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     4585..4604
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4823..4891,1..36))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
       61 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
      121 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
      181 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
      241 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
      301 actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac
      361 cttatgtatc atacacatac gatttaggtg acactataga acgcggccgc cagctgaagc
      421 ttcgtacgct gcaggtcgac ggatcggtga cggtgctggt ttaattaaca tggtgtccaa
      481 aggagaggag ttaatcaagg aaaacatgag aatgaaagtt gtcatggagg gctccgttaa
      541 tggtcaccaa ttcaagtgta caggggaagg tgaaggtaat ccttacatgg gtacacaaac
      601 tatgagaatt aaagtaattg aaggcggacc actaccattt gcatttgaca ttctggcaac
      661 gtcattcatg tacggatcac gaactttcat caagtaccct aaaggtatac cagacttttt
      721 caagcaatct tttccagagg gttttacatg ggaaagggtt acaagatacg aagatggggg
      781 tgtcgtcaca gttatgcaag atacttcatt agaagatggc tgccttgtct atcatgtgca
      841 agtaagaggg gtgaattttc cttctaacgg acctgtgatg cagaaaaaga ccaaaggttg
      901 ggaaccaaat actgaaatga tgtacccagc tgatggaggt ttgagaggct acacacacat
      961 ggcgcttaaa gttgatggtg gaggtcattt gtcttgtagt tttgttacca cttatcgttc
     1021 taaaaagact gttggcaata tcaaaatgcc aggaatacat gctgtagacc acagactaga
     1081 aagactcgaa gagagcgata acgaaatgtt cgttgtacag agagagcatg ccgtagccaa
     1141 atttgctggc ttaggcggtg gtatggatga attgtataag taaggcgcgc cacttctaaa
     1201 taagcgaatt tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt
     1261 gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac
     1321 tctttcctgt aggtcaggtt gctttctcag gtatagtatg aggtcgctct tattgaccac
     1381 acctctaccg gcagatccgc tagggataac agggtaatat agatctgttt agcttgcctc
     1441 gtccccgccg ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct
     1501 tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc
     1561 ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta
     1621 cggctcctcg ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg
     1681 tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt
     1741 ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca
     1801 taaacaacca tgggtaagga aaagactcac gtttcgaggc cgcgattaaa ttccaacatg
     1861 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     1921 atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     1981 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     2041 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     2101 gcgatccccg gcaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     2161 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     2221 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     2281 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     2341 aaagaaatgc ataagctttt gccattctca ccggattcag tcgtcactca tggtgatttc
     2401 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     2461 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     2521 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2581 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagtactgac aataaaaaga
     2641 ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt
     2701 aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga
     2761 agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac
     2821 tgctgtcgat tcgatactaa cgccgccatc cagtttaaac gagctcgaat tcatcgatga
     2881 tatcagatcc actagtggcc tatgcggccg cggatctgcc ggtctcccta tagtgagtcg
     2941 tattaatttc gataagccag gttaacctgc attaatgaat cggccaacgc gcggggagag
     3001 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3061 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3121 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3181 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3241 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3301 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3361 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     3421 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3481 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3541 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3601 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     3661 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3721 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3781 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3841 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3901 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3961 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4021 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4081 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4141 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4201 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4261 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4321 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4381 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4441 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4501 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4561 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4621 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4681 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4741 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4801 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4861 gttattgtct catgagcgga tacatatttg a
//
