LOCUS       sequence-271019-        4437 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 4437)
  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 4437)
  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:hphMX4
FEATURES             Location/Qualifiers
     source          1..4437
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            41..1616
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        41..384
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             385..1413
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             385..1413
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      1419..1616
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1723..1742)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1724..1742)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1829..1846)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2000..2588)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2080..2099)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2759..3550,3551..3619))
                     /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     3382..3401
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3620..3724)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3792..3810
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3848..3870)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3970..3989
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4070..4088
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4070..4087
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4169..4192
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      4216..4403
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4271..4295)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
ORIGIN
        1 tgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac
       61 cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat
      121 ttagcccata catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc
      181 gcgaagcaaa aattacggct cctcgctgcg gacctgcgag cagggaaacg ctcccctcac
      241 agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt
      301 gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca
      361 catcacatcc gaacataaac aaccatgggt aaaaagcctg aactcaccgc gacgtctgtc
      421 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
      481 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
      541 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg
      601 ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc
      661 tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt
      721 ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc
      781 gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata
      841 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt
      901 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc
      961 cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata
     1021 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac
     1081 atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg
     1141 aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt
     1201 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     1261 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     1321 agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga
     1381 cgccccagca ctcgtccgag ggcaaaggaa taatcagtac tgacaataaa aagattcttg
     1441 ttttcaagaa cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa
     1501 atgttagcgt gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa
     1561 gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt
     1621 cgattcgata ctaacgccgc catccagtgt cgaaaacgag ctcgaattca tcgatgatat
     1681 cagatccact agtggcctat gcggccgcgg atctgccggt ctccctatag tgagtcgtat
     1741 taatttcgat aagccaggtt aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1801 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1861 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1921 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1981 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2041 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2101 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2161 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2221 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2281 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2341 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2401 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2461 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2521 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2581 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2641 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2701 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2761 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2821 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     2881 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     2941 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3001 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3061 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3121 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3181 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3241 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3301 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3361 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3421 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3481 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3541 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3601 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3661 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3721 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     3781 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     3841 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     3901 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     3961 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatggac atattgtcgt
     4021 tagaacgcgg ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac
     4081 actatagaac gcggccgcca gctgaagctt cgtacgctgc aggtcgacgg atccccgggt
     4141 taattaactg cccgaaaccg gtggtggcga ttacaaggac gacgatgaca agtaaggcgc
     4201 gccacttcta aataagcgaa tttcttatga tttatgattt ttattattaa ataagttata
     4261 aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta
     4321 ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagta tgaggtcgct
     4381 cttattgacc acacctctac cggcagatcc gctagggata acagggtaat atagatc
//
