LOCUS       sequence-271060-        4117 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 4117)
  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 4117)
  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:HIS3MX6
FEATURES             Location/Qualifiers
     source          1..4117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      26..213
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(81..105)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        288..631
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             632..1330
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1334..1531
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1635..1654)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1636..1654)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1741..1758)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1912..2500)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1992..2011)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2671..3462,3463..3531))
                     /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     3294..3313
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3532..3636)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3704..3722
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3760..3782)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3882..3901
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3982..4000
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3982..3999
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             join(4096..4117,1..2)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 agtaaggcgc gccacttcta aataagcgaa tttcttatga tttatgattt ttattattaa
       61 ataagttata aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg
      121 aaaattctta ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagta
      181 tgaggtcgct cttattgacc acacctctac cggcagatcc gctagggata acagggtaat
      241 atagatctgt ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc
      301 agaataccct ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc
      361 cgtacattta gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc
      421 gcacggcgcg aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc
      481 ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt
      541 aggatttgcc actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca
      601 gttctcacat cacatccgaa cataaacaac catggcagaa ccagcccaaa aaaagcaaaa
      661 acaaactgtt caggagcgca aggcgtttat ctcccgtatc actaatgaaa ctaaaattca
      721 aatcgctatt tcgctgaatg gtggttatat tcaaataaaa gattcgattc ttcctgcaaa
      781 gaaggatgac gatgtagctt cccaagctac tcagtcacag gtcatcgata ttcacacagg
      841 tgttggcttt ttggatcata tgatccatgc gttggcaaaa cactctggtt ggtctcttat
      901 tgttgaatgt attggtgacc tgcacattga cgatcaccat actaccgaag attgcggtat
      961 cgcattaggg caagcgttca aagaagcaat gggtgctgtc cgtggtgtaa aaagattcgg
     1021 tactgggttc gcaccattgg atgaggcgct atcacgtgcc gtagtcgatt tatctagtag
     1081 accatttgct gtaatcgacc ttggattgaa gagagagatg attggtgatt tatccactga
     1141 aatgattcca cactttttgg aaagtttcgc ggaggcggcc agaattactt tgcatgttga
     1201 ttgtctgaga ggtttcaacg atcaccacag aagtgagagt gcgttcaagg ctttggctgt
     1261 tgccataaga gaagctattt ctagcaatgg caccaatgac gttccctcaa ccaaaggtgt
     1321 tttgatgtga agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata
     1381 gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt
     1441 tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt
     1501 caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc
     1561 agtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct atgcggccgc
     1621 ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca
     1681 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1741 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1801 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1861 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1921 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1981 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2041 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2101 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2161 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2221 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2281 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2341 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2401 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2461 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2521 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2581 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2641 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2701 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2761 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2821 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2881 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2941 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3001 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3061 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3121 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3181 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3241 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3301 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3361 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3421 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3481 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3541 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3601 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3661 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     3721 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     3781 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     3841 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     3901 actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac
     3961 cttatgtatc atacacatac gatttaggtg acactataga acgcggccgc cagctgaagc
     4021 ttcgtacgct gcaggtcgac ggatccccgg gttaattaag gggatagctt atcctggtta
     4081 cttagactgt taaacgatta caaggacgac gatgaca
//
