LOCUS       sequence-271136-        4102 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 4102)
  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 4102)
  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:HIS3MX6
FEATURES             Location/Qualifiers
     source          1..4102
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     70..89
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        170..188
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     170..187
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             269..292
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      316..503
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(371..395)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        578..921
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             922..1620
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1624..1821
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1925..1944)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1926..1944)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2031..2048)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2202..2790)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2282..2301)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2961..3752,3753..3821))
                     /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     3584..3603
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3822..3926)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3994..4012
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4050..4072)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
       61 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatggac atattgtcgt
      121 tagaacgcgg ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac
      181 actatagaac gcggccgcca gctgaagctt cgtacgctgc aggtcgacgg atccccgggt
      241 taattaactg cccgaaaccg gtggtggcga ttacaaggac gacgatgaca agtaaggcgc
      301 gccacttcta aataagcgaa tttcttatga tttatgattt ttattattaa ataagttata
      361 aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta
      421 ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagta tgaggtcgct
      481 cttattgacc acacctctac cggcagatcc gctagggata acagggtaat atagatctgt
      541 ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct
      601 ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta
      661 gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg
      721 aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga
      781 cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc
      841 actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat
      901 cacatccgaa cataaacaac catggcagaa ccagcccaaa aaaagcaaaa acaaactgtt
      961 caggagcgca aggcgtttat ctcccgtatc actaatgaaa ctaaaattca aatcgctatt
     1021 tcgctgaatg gtggttatat tcaaataaaa gattcgattc ttcctgcaaa gaaggatgac
     1081 gatgtagctt cccaagctac tcagtcacag gtcatcgata ttcacacagg tgttggcttt
     1141 ttggatcata tgatccatgc gttggcaaaa cactctggtt ggtctcttat tgttgaatgt
     1201 attggtgacc tgcacattga cgatcaccat actaccgaag attgcggtat cgcattaggg
     1261 caagcgttca aagaagcaat gggtgctgtc cgtggtgtaa aaagattcgg tactgggttc
     1321 gcaccattgg atgaggcgct atcacgtgcc gtagtcgatt tatctagtag accatttgct
     1381 gtaatcgacc ttggattgaa gagagagatg attggtgatt tatccactga aatgattcca
     1441 cactttttgg aaagtttcgc ggaggcggcc agaattactt tgcatgttga ttgtctgaga
     1501 ggtttcaacg atcaccacag aagtgagagt gcgttcaagg ctttggctgt tgccataaga
     1561 gaagctattt ctagcaatgg caccaatgac gttccctcaa ccaaaggtgt tttgatgtga
     1621 agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat
     1681 attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac
     1741 atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg
     1801 tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc agtttaaacg
     1861 agctcgaatt catcgatgat atcagatcca ctagtggcct atgcggccgc ggatctgccg
     1921 gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca ttaatgaatc
     1981 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2041 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2101 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2161 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2221 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2281 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2341 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2401 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2461 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2521 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2581 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2641 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2701 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2761 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2821 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2881 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2941 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3001 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3061 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3121 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3181 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3241 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3301 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3361 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3421 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3481 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3541 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3601 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3661 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3721 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3781 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3841 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3901 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     3961 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     4021 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     4081 acagcttgtc tgtaagcgga tg
//
