LOCUS       sequence-271038-        4215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR-based C-terminal SRS-FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4215)
  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 4215)
  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-SRS-FLAG:kanMX6
FEATURES             Location/Qualifiers
     source          1..4215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..46
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        127..145
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     127..144
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             226..249
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      273..460
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(328..352)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            535..1891
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        535..878
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             879..1688
                     /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(946..965)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1694..1891
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1995..2014)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1996..2014)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2101..2118)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2272..2860)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2352..2371)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3031..3822,3823..3891))
                     /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     3654..3673
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3892..3996)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4064..4082
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4120..4142)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
       61 tatggacata ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca
      121 catacgattt aggtgacact atagaacgcg gccgccagct gaagcttcgt acgctgcagg
      181 tcgacggatc cccgggttaa ttaactgccc gaaaccggtg gtggcgatta caaggacgac
      241 gatgacaagt aaggcgcgcc acttctaaat aagcgaattt cttatgattt atgattttta
      301 ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt
      361 taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg
      421 tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct agggataaca
      481 gggtaatata gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg
      541 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac
      601 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt
      661 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga
      721 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata
      781 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta
      841 ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaaggaa aagactcacg
      901 tttcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
      961 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     1021 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     1081 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     1141 ctcctgatga tgcatggtta ctcaccactg cgatccccgg caaaacagca ttccaggtat
     1201 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     1261 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     1321 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     1381 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac
     1441 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     1501 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     1561 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     1621 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     1681 ttttctaatc agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata
     1741 gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt
     1801 tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt
     1861 caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc
     1921 agtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct atgcggccgc
     1981 ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca
     2041 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2101 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2161 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2221 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2281 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2341 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2401 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2461 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2521 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2581 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2641 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2701 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2761 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2821 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2881 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2941 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3001 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3061 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3121 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3181 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3241 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3301 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3361 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3421 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3481 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3541 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3601 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3661 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3721 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3781 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3841 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3901 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3961 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4021 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     4081 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     4141 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     4201 gtcagcgggt gttgg
//
