LOCUS       sequence-9658-ex        4234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4234)
  AUTHORS   Funakoshi M, Hochstrasser M
  TITLE     Small epitope-linker modules for PCR-based C-terminal tagging in 
            Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2009 Feb 25. 26(3):185-192.
  PUBMED    19243080
REFERENCE   2  (bases 1 to 4234)
  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. 2009
            Feb 25. 26(3):185-192."
COMMENT     Sequence Label: pFA6a-6xGLY-S-tag-kanMX6
FEATURES             Location/Qualifiers
     source          1..4234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..123
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      147..334
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(202..226)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            409..1765
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        409..752
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             753..1562
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(820..839)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1568..1765
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1869..1888)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1870..1888)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1975..1992)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2146..2734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2226..2245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2905..3696,3697..3765))
                     /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     3528..3547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3766..3870)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3938..3956
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3994..4016)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4116..4135
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4216..4234
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4216..4233
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaaa agaaaccgct gctgctaaat tcgaacgcca gcacatggac
      121 agctagggcg cgccacttct aaataagcga atttcttatg atttatgatt tttattatta
      181 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
      241 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt
      301 atgaggtcgc tcttattgac cacacctcta ccggcagatc cgctagggat aacagggtaa
      361 tatagatctg tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc
      421 cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc
      481 ccgtacattt agcccataca tccccatgta taatcatttg catccataca ttttgatggc
      541 cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca gggaaacgct
      601 cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt
      661 taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac
      721 agttctcaca tcacatccga acataaacaa ccatgggtaa ggaaaagact cacgtttcga
      781 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
      841 atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt
      901 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
      961 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     1021 atgatgcatg gttactcacc actgcgatcc ccggcaaaac agcattccag gtattagaag
     1081 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     1141 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     1201 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     1261 gctggcctgt tgaacaagtc tggaaagaaa tgcataagct tttgccattc tcaccggatt
     1321 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     1381 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     1441 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     1501 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     1561 aatcagtact gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt
     1621 ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct
     1681 cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg
     1741 tatgtgaatg ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccagttta
     1801 aacgagctcg aattcatcga tgatatcaga tccactagtg gcctatgcgg ccgcggatct
     1861 gccggtctcc ctatagtgag tcgtattaat ttcgataagc caggttaacc tgcattaatg
     1921 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1981 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2041 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2101 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2161 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2221 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2281 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2341 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2401 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2461 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2521 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2581 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2641 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2701 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2761 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2821 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2881 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2941 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3001 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3061 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3121 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3181 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3241 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3301 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3361 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3421 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3481 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3541 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3601 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3661 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3721 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     3781 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3841 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     3901 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     3961 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     4021 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     4081 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     4141 agtgcaccat atggacatat tgtcgttaga acgcggctac aattaataca taaccttatg
     4201 tatcatacac atacgattta ggtgacacta taga
//
