LOCUS       sequence-271057-        4230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR-based C-terminal S6-FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4230)
  AUTHORS   Nemec AA, Tomko RJ Jr
  TITLE     A suite of PCR-based peptide tagging plasmids to permit 
            epitope-targeted enzymatic functionalization of yeast proteins.
  JOURNAL   Yeast. 2020 May 13. doi: 10.1002/yea.3471.
  PUBMED    32401365
REFERENCE   2  (bases 1 to 4230)
  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: "Yeast. 2020
            May 13. doi: 10.1002/yea.3471."
COMMENT     Sequence Label: pFA6a-S6-FLAG:kanMX6
FEATURES             Location/Qualifiers
     source          1..4230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        206..224
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     206..223
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             320..343
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      367..554
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(422..446)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            629..1985
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        629..972
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             973..1782
                     /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(1040..1059)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1788..1985
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2089..2108)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2090..2108)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2195..2212)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2366..2954)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2446..2465)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3125..3916,3917..3985))
                     /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     3748..3767
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3986..4090)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4158..4176
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(4214..4230,1..6))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
       61 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
      121 ttgtactgag agtgcaccat atggacatat tgtcgttaga acgcggctac aattaataca
      181 taaccttatg tatcatacac atacgattta ggtgacacta tagaacgcgg ccgccagctg
      241 aagcttcgta cgctgcaggt cgacggatcc ccgggttaat taaggggata gcttatcctg
      301 gttacttaga ctgttaaacg attacaagga cgacgatgac aagtaaggcg cgccacttct
      361 aaataagcga atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata
      421 agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt
      481 aactctttcc tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac
      541 cacacctcta ccggcagatc cgctagggat aacagggtaa tatagatctg tttagcttgc
      601 ctcgtccccg ccgggtcacc cggccagcga catggaggcc cagaataccc tccttgacag
      661 tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca
      721 tccccatgta taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa
      781 ttacggctcc tcgctgcaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa
      841 ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt
      901 cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga
      961 acataaacaa ccatgggtaa ggaaaagact cacgtttcga ggccgcgatt aaattccaac
     1021 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     1081 acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     1141 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     1201 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     1261 actgcgatcc ccggcaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     1321 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     1381 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     1441 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     1501 tggaaagaaa tgcataagct tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     1561 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     1621 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     1681 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     1741 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagtact gacaataaaa
     1801 agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat
     1861 tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg
     1921 cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta
     1981 tactgctgtc gattcgatac taacgccgcc atccagttta aacgagctcg aattcatcga
     2041 tgatatcaga tccactagtg gcctatgcgg ccgcggatct gccggtctcc ctatagtgag
     2101 tcgtattaat ttcgataagc caggttaacc tgcattaatg aatcggccaa cgcgcgggga
     2161 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2221 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2281 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2341 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2401 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2461 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2521 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2581 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2641 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2701 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2761 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2821 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2881 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2941 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3001 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3061 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3121 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3181 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3241 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3301 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3361 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3421 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3481 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     3541 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     3601 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3661 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     3721 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     3781 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     3841 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     3901 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     3961 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4021 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4081 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     4141 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     4201 atgacggtga aaacctctga cacatgcagc
//
