LOCUS       sequence-197818-        4873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast optimized 
            mTagBFP2 with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4873)
  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 4873)
  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-yomTagBFP2-Kan
FEATURES             Location/Qualifiers
     source          1..4873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        52..70
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     52..69
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      856..1043
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(911..935)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1118..2474
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1118..1461
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1462..2271
                     /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(1529..1548)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2277..2474
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2578..2597)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2579..2597)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2684..2701)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2855..3443)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2935..2954)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3614..4405,4406..4474))
                     /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     4237..4256
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4475..4579)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4647..4665
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4703..4725)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4825..4844
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg
       61 acactataga acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatcggtga
      121 cggtgctggt ttaattaaca tgagtgagtt gattaaggaa aacatgcaca tgaagttgta
      181 catggaagga acagtagata atcatcattt caaatgtacg tcagaaggtg agggcaaacc
      241 ttatgaagga actcaaacta tgagaattaa agtcgttgag ggtggcccac taccatttgc
      301 attcgatatc ctcgcaacat cttttctgta tggttctaaa acctttatca accatacaca
      361 aggcatacca gatttcttta agcaatcatt tcctgaaggt ttcacatggg aaagagtaac
      421 tacctatgaa gatggtggtg tccttacagc tactcaagac acgtctttgc aggatggttg
      481 tttaatctac aatgttaaga taagaggagt gaactttaca tccaatggac cagtcatgca
      541 aaagaaaact ctaggttggg aggcattcac agaaactttg tacccagccg atgggggctt
      601 agaagggaga aatgacatgg ctttaaaact ggttggcggt agccatttga ttgcgaacgc
      661 taaaacaacc tacagaagca aaaagccagc taaaaatctt aagatgcctg tatactacgt
      721 ggattacaga ttagaaagga aggaagctaa taatgaaact tacgttgaac agcacgaggt
      781 ggccgttgcc cgttattgcg accttccttc aaaattggga cacaaactaa actaaggcgc
      841 gccacttcta aataagcgaa tttcttatga tttatgattt ttattattaa ataagttata
      901 aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta
      961 ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagta tgaggtcgct
     1021 cttattgacc acacctctac cggcagatcc gctagggata acagggtaat atagatctgt
     1081 ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct
     1141 ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta
     1201 gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg
     1261 aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga
     1321 cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc
     1381 actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat
     1441 cacatccgaa cataaacaac catgggtaag gaaaagactc acgtttcgag gccgcgatta
     1501 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     1561 tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa
     1621 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     1681 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     1741 ttactcacca ctgcgatccc cggcaaaaca gcattccagg tattagaaga atatcctgat
     1801 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     1861 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     1921 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     1981 gaacaagtct ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact
     2041 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     2101 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     2161 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     2221 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagtactg
     2281 acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag
     2341 ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct
     2401 gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc
     2461 tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtttaa acgagctcga
     2521 attcatcgat gatatcagat ccactagtgg cctatgcggc cgcggatctg ccggtctccc
     2581 tatagtgagt cgtattaatt tcgataagcc aggttaacct gcattaatga atcggccaac
     2641 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2701 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2761 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2821 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2881 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2941 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3001 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3061 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3121 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3181 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3241 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3301 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3361 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3421 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3481 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3541 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3601 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3661 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3721 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3781 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3841 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3901 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3961 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4021 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4081 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4141 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4201 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4261 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4321 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4381 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4441 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4501 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4561 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     4621 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     4681 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     4741 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     4801 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     4861 tggacatatt gtc
//
