LOCUS       sequence-271013-        3972 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 3972)
  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 3972)
  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:natMX4
FEATURES             Location/Qualifiers
     source          1..3972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3..25)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     125..144
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        225..243
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     225..242
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      362..549
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(417..441)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            624..1743
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        624..967
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             968..1540
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1546..1743
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1850..1869)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1851..1869)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1956..1973)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2127..2715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2207..2226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2886..3677,3678..3746))
                     /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     3509..3528
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3747..3851)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3919..3937
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
       61 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
      121 tatgcggcat cagagcagat tgtactgaga gtgcaccata tggacatatt gtcgttagaa
      181 cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat
      241 agaacgcggc cgccagctga agcttcgtac gctgcaggtc gacggatccc cgggttaatt
      301 aaggggatag cctggacatg ctcgagtggt ctctgatgta aggcgcgcca cttctaaata
      361 agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt
      421 atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc
      481 tttcctgtag gtcaggttgc tttctcaggt atagtatgag gtcgctctta ttgaccacac
      541 ctctaccggc agatccgcta gggataacag ggtaatatag atctgtttag cttgcctcgt
      601 ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg
      661 acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc
      721 atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg
      781 gctcctcgct gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc
      841 cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct
      901 ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata
      961 aacaaccatg ggtaccactc ttgacgacac ggcttaccgg taccgcacca gtgtcccggg
     1021 ggacgccgag gccatcgagg cactggatgg gtccttcacc accgacaccg tcttccgcgt
     1081 caccgccacc ggggacggct tcaccctgcg ggaggtgccg gtggacccgc ccctgaccaa
     1141 ggtgttcccc gacgacgaat cggacgacga atcggacgac ggggaggacg gcgacccgga
     1201 ctcccggacg ttcgtcgcgt acggggacga cggcgacctg gcgggcttcg tggtcgtctc
     1261 gtactccggc tggaaccgcc ggctgaccgt cgaggacatc gaggtcgccc cggagcaccg
     1321 ggggcacggg gtcgggcgcg cgttgatggg gctcgcgacg gagttcgccc gcgagcgggg
     1381 cgccgggcac ctctggctgg aggtcaccaa cgtcaacgca ccggcgatcc acgcgtaccg
     1441 gcggatgggg ttcaccctct gcggcctgga caccgccctg tacgacggca ccgcctcgga
     1501 cggcgagcag gcgctctaca tgagcatgcc ctgcccctaa tcagtactga caataaaaag
     1561 attcttgttt tcaagaactt gtcatttgta tagttttttt atattgtagt tgttctattt
     1621 taatcaaatg ttagcgtgat ttatattttt tttcgcctcg acatcatctg cccagatgcg
     1681 aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata
     1741 ctgctgtcga ttcgatacta acgccgccat ccagtgtcga aaacgagctc gaattcatcg
     1801 atgatatcag atccactagt ggcctatgcg gccgcggatc tgccggtctc cctatagtga
     1861 gtcgtattaa tttcgataag ccaggttaac ctgcattaat gaatcggcca acgcgcgggg
     1921 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     1981 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2041 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2101 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2161 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2221 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2281 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2341 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2401 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2461 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2521 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2581 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2641 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2701 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2761 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2821 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2881 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2941 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3001 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3061 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3121 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3181 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3241 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3301 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3361 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3421 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3481 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3541 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3601 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3661 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3721 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3781 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3841 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     3901 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     3961 gatgacggtg aa
//
