LOCUS       sequence-181116-        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     complement(40..62)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     162..181
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        262..280
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     262..279
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             350..1060
                     /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      1084..1271
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1139..1163)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1346..2702
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1346..1689
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1690..2499
                     /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(1757..1776)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2505..2702
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2806..2825)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2807..2825)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2912..2929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3083..3671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3163..3182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3842..4633,4634..4702))
                     /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     4465..4484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4703..4807)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(4875..4891,1..2)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
       61 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
      121 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
      181 actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac
      241 cttatgtatc atacacatac gatttaggtg acactataga acgcggccgc cagctgaagc
      301 ttcgtacgct gcaggtcgac ggatcggtga cggtgctggt ttaattaaca tggtgtccaa
      361 aggagaggag ttaatcaagg aaaacatgag aatgaaagtt gtcatggagg gctccgttaa
      421 tggtcaccaa ttcaagtgta caggggaagg tgaaggtaat ccttacatgg gtacacaaac
      481 tatgagaatt aaagtaattg aaggcggacc actaccattt gcatttgaca ttctggcaac
      541 gtcattcatg tacggatcac gaactttcat caagtaccct aaaggtatac cagacttttt
      601 caagcaatct tttccagagg gttttacatg ggaaagggtt acaagatacg aagatggggg
      661 tgtcgtcaca gttatgcaag atacttcatt agaagatggc tgccttgtct atcatgtgca
      721 agtaagaggg gtgaattttc cttctaacgg acctgtgatg cagaaaaaga ccaaaggttg
      781 ggaaccaaat actgaaatga tgtacccagc tgatggaggt ttgagaggct acacacacat
      841 ggcgcttaaa gttgatggtg gaggtcattt gtcttgtagt tttgttacca cttatcgttc
      901 taaaaagact gttggcaata tcaaaatgcc aggaatacat gctgtagacc acagactaga
      961 aagactcgaa gagagcgata acgaaatgtt cgttgtacag agagagcatg ccgtagccaa
     1021 atttgctggc ttaggcggtg gtatggatga attgtataag taaggcgcgc cacttctaaa
     1081 taagcgaatt tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt
     1141 gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac
     1201 tctttcctgt aggtcaggtt gctttctcag gtatagtatg aggtcgctct tattgaccac
     1261 acctctaccg gcagatccgc tagggataac agggtaatat agatctgttt agcttgcctc
     1321 gtccccgccg ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct
     1381 tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc
     1441 ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta
     1501 cggctcctcg ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg
     1561 tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt
     1621 ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca
     1681 taaacaacca tgggtaagga aaagactcac gtttcgaggc cgcgattaaa ttccaacatg
     1741 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     1801 atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     1861 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     1921 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     1981 gcgatccccg gcaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     2041 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     2101 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     2161 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     2221 aaagaaatgc ataagctttt gccattctca ccggattcag tcgtcactca tggtgatttc
     2281 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     2341 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     2401 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2461 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagtactgac aataaaaaga
     2521 ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt
     2581 aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga
     2641 agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac
     2701 tgctgtcgat tcgatactaa cgccgccatc cagtttaaac gagctcgaat tcatcgatga
     2761 tatcagatcc actagtggcc tatgcggccg cggatctgcc ggtctcccta tagtgagtcg
     2821 tattaatttc gataagccag gttaacctgc attaatgaat cggccaacgc gcggggagag
     2881 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2941 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3001 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3061 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3121 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3181 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3241 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3301 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3361 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3421 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3481 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3541 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3601 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3661 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3721 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3781 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3841 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3901 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3961 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4021 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4081 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4141 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4201 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4261 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4321 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4381 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4441 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4501 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4561 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4621 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4681 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4741 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4801 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     4861 cattaaccta taaaaatagg cgtatcacga g
//
