LOCUS       sequence-62222-e        4888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            photoactivatable mCherry with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4888)
  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 4888)
  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-yoPA-mCherry-Kan
FEATURES             Location/Qualifiers
     source          1..4888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     109..128
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        209..227
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     209..226
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      1028..1215
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1083..1107)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1290..2646
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1290..1633
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1634..2443
                     /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(1701..1720)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2449..2646
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2750..2769)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2751..2769)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2856..2873)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3027..3615)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3107..3126)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3786..4577,4578..4646))
                     /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     4409..4428
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4647..4751)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4819..4837
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(4875..4888,1..9))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
       61 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
      121 agattgtact gagagtgcac catatggaca tattgtcgtt agaacgcggc tacaattaat
      181 acataacctt atgtatcata cacatacgat ttaggtgaca ctatagaacg cggccgccag
      241 ctgaagcttc gtacgctgca ggtcgacgga tcggtgacgg tgctggttta attaacatgg
      301 tcagtaaagg ggaggaagat aatatggcaa tcattaaaga gtttatgcgt tttaaagttc
      361 acatggaggg ttcagttaac ggccatgttt ttgaaattga aggtgaagga gaaggaaggc
      421 catacgaagg tactcaaaca gcaaagctga aagtaaccaa aggcggtcca ttaccattca
      481 cttgggatat cctttctcct caattcatgt acggttccaa tgcctacgtg aagcaccctg
      541 ctgatatccc agactacttc aaactatcat ttcctgaagg ctttaagtgg gaaagagtca
      601 tgaagttcga agatggcggt gtggtcactg ttactcaaga ctcttcctta caagatggcg
      661 aattcatata caaagttaag ctaagaggaa caaattttcc atcagatggg cctgtaatgc
      721 agaaaaagac catggggtgg gaggccttat ctgagagaat gtatccagaa gatggagcgc
      781 tcaaaggtga ggttaaacct agagtaaaat tgaaagacgg tggtcattac gatgctgaag
      841 taaagacaac ttacaaggct aaaaagccag ttcaattgcc aggagcatat aacgtcaaca
      901 gaaaacttga cattacaagt cataatgaag attacacaat agtggaacag tatgaaagag
      961 ctgaaggtag acattctacg ggtggaatgg acgagttgta taagtaaggc gcgccacttc
     1021 taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta taaaaaaaat
     1081 aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct tattcttgag
     1141 taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg ctcttattga
     1201 ccacacctct accggcagat ccgctaggga taacagggta atatagatct gtttagcttg
     1261 cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc ctccttgaca
     1321 gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt tagcccatac
     1381 atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg cgaagcaaaa
     1441 attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca gacgcgttga
     1501 attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg ccactgaggt
     1561 tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac atcacatccg
     1621 aacataaaca accatgggta aggaaaagac tcacgtttcg aggccgcgat taaattccaa
     1681 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     1741 gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     1801 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     1861 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     1921 cactgcgatc cccggcaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     1981 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     2041 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     2101 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     2161 ctggaaagaa atgcataagc ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     2221 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     2281 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     2341 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     2401 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagtac tgacaataaa
     2461 aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt agttgttcta
     2521 ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat ctgcccagat
     2581 gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct
     2641 atactgctgt cgattcgata ctaacgccgc catccagttt aaacgagctc gaattcatcg
     2701 atgatatcag atccactagt ggcctatgcg gccgcggatc tgccggtctc cctatagtga
     2761 gtcgtattaa tttcgataag ccaggttaac ctgcattaat gaatcggcca acgcgcgggg
     2821 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2881 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2941 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3001 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3061 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3121 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3181 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     3241 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3301 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3361 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3421 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     3481 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3541 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3601 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3661 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3721 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3781 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3841 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3901 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3961 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4021 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4081 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4141 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4201 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4261 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4321 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4381 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4441 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4501 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4561 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4621 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4681 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4741 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     4801 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     4861 gatgacggtg aaaacctctg acacatgc
//
