LOCUS       sequence-271059-        4093 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 4093)
  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 4093)
  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:HIS3MX6
FEATURES             Location/Qualifiers
     source          1..4093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     96..115
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        196..214
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     196..213
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      333..520
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(388..412)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        595..938
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             939..1637
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1641..1838
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1942..1961)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1943..1961)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2048..2065)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2219..2807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2299..2318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2978..3769,3770..3838))
                     /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     3601..3620
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3839..3943)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4011..4029
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4067..4089)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
       61 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
      121 agtgcaccat atggacatat tgtcgttaga acgcggctac aattaataca taaccttatg
      181 tatcatacac atacgattta ggtgacacta tagaacgcgg ccgccagctg aagcttcgta
      241 cgctgcaggt cgacggatcc ccgggttaat taaggggata gcctggacat gctcgagtgg
      301 tctctgatgt aaggcgcgcc acttctaaat aagcgaattt cttatgattt atgattttta
      361 ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt
      421 taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg
      481 tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct agggataaca
      541 gggtaatata gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg
      601 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac
      661 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt
      721 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga
      781 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata
      841 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta
      901 ggatacagtt ctcacatcac atccgaacat aaacaaccat ggcagaacca gcccaaaaaa
      961 agcaaaaaca aactgttcag gagcgcaagg cgtttatctc ccgtatcact aatgaaacta
     1021 aaattcaaat cgctatttcg ctgaatggtg gttatattca aataaaagat tcgattcttc
     1081 ctgcaaagaa ggatgacgat gtagcttccc aagctactca gtcacaggtc atcgatattc
     1141 acacaggtgt tggctttttg gatcatatga tccatgcgtt ggcaaaacac tctggttggt
     1201 ctcttattgt tgaatgtatt ggtgacctgc acattgacga tcaccatact accgaagatt
     1261 gcggtatcgc attagggcaa gcgttcaaag aagcaatggg tgctgtccgt ggtgtaaaaa
     1321 gattcggtac tgggttcgca ccattggatg aggcgctatc acgtgccgta gtcgatttat
     1381 ctagtagacc atttgctgta atcgaccttg gattgaagag agagatgatt ggtgatttat
     1441 ccactgaaat gattccacac tttttggaaa gtttcgcgga ggcggccaga attactttgc
     1501 atgttgattg tctgagaggt ttcaacgatc accacagaag tgagagtgcg ttcaaggctt
     1561 tggctgttgc cataagagaa gctatttcta gcaatggcac caatgacgtt ccctcaacca
     1621 aaggtgtttt gatgtgaagt actgacaata aaaagattct tgttttcaag aacttgtcat
     1681 ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata
     1741 ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat
     1801 catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc
     1861 gccatccagt ttaaacgagc tcgaattcat cgatgatatc agatccacta gtggcctatg
     1921 cggccgcgga tctgccggtc tccctatagt gagtcgtatt aatttcgata agccaggtta
     1981 acctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2041 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2101 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2161 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2221 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2281 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2341 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2401 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2461 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2521 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2581 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2641 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2701 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2761 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2821 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2881 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2941 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3001 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3061 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3121 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3181 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3241 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3301 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3361 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3421 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3481 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3541 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3601 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3661 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3721 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3781 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3841 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3901 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3961 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     4021 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     4081 agctcccgga gac
//
