LOCUS       sequence-271063-        4428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR-based C-terminal A1 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4428)
  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 4428)
  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-A1:hphMX4
FEATURES             Location/Qualifiers
     source          1..4428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..43
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        124..142
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     124..141
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      261..448
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(316..340)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            523..2098
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        523..866
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             867..1895
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             867..1895
                     /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      1901..2098
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2205..2224)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2206..2224)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2311..2328)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2482..3070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2562..2581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3241..4032,4033..4101))
                     /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     3864..3883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4102..4206)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4274..4292
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4330..4352)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
       61 ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat
      121 acgatttagg tgacactata gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg
      181 acggatcccc gggttaatta aggggatagc ctggacatgc tcgagtggtc tctgatgtaa
      241 ggcgcgccac ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag
      301 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
      361 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg
      421 tcgctcttat tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga
      481 tctgtttagc ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat
      541 accctccttg acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac
      601 atttagccca tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg
      661 gcgcgaagca aaaattacgg ctcctcgctg cggacctgcg agcagggaaa cgctcccctc
      721 acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat
      781 ttgccactga ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct
      841 cacatcacat ccgaacataa acaaccatgg gtaaaaagcc tgaactcacc gcgacgtctg
      901 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
      961 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
     1021 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg
     1081 cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg acctattgca
     1141 tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
     1201 ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga
     1261 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
     1321 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     1381 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     1441 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
     1501 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
     1561 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
     1621 ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc
     1681 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
     1741 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
     1801 gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
     1861 gacgccccag cactcgtccg agggcaaagg aataatcagt actgacaata aaaagattct
     1921 tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc tattttaatc
     1981 aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag atgcgaagtt
     2041 aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg ctatactgct
     2101 gtcgattcga tactaacgcc gccatccagt gtcgaaaacg agctcgaatt catcgatgat
     2161 atcagatcca ctagtggcct atgcggccgc ggatctgccg gtctccctat agtgagtcgt
     2221 attaatttcg ataagccagg ttaacctgca ttaatgaatc ggccaacgcg cggggagagg
     2281 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2341 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2401 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2461 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2521 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2581 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2641 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2701 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2761 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2821 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2881 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2941 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3001 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3061 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3121 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3181 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3241 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3301 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3361 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3421 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3481 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3541 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3601 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3661 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3721 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3781 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3841 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3901 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3961 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4021 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4081 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4141 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4201 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4261 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     4321 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     4381 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgg
//
