LOCUS       sequence-9646-ex        4255 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 4255)
  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 4255)
  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-3xFLAG-kanMX6
FEATURES             Location/Qualifiers
     source          1..4255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..144
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      168..355
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(223..247)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            430..1786
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        430..773
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             774..1583
                     /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(841..860)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1589..1786
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1890..1909)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1891..1909)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1996..2013)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2167..2755)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2247..2266)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2926..3717,3718..3786))
                     /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     3549..3568
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3787..3891)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3959..3977
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4015..4037)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4137..4156
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4237..4255
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4237..4254
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaga ctacaaagac catgacggtg attataaaga tcatgacatc
      121 gactacaagg atgacgatga caagtagggc gcgccacttc taaataagcg aatttcttat
      181 gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa
      241 gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca
      301 ggttgctttc tcaggtatag tatgaggtcg ctcttattga ccacacctct accggcagat
      361 ccgctaggga taacagggta atatagatct gtttagcttg cctcgtcccc gccgggtcac
      421 ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt gcgcagctca
      481 ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt ataatcattt
      541 gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag
      601 acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc
      661 ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct
      721 tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca accatgggta
      781 aggaaaagac tcacgtttcg aggccgcgat taaattccaa catggatgct gatttatatg
      841 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg
      901 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
      961 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     1021 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggcaaaa
     1081 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     1141 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     1201 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     1261 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc
     1321 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     1381 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     1441 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     1501 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     1561 tgatgctcga tgagtttttc taatcagtac tgacaataaa aagattcttg ttttcaagaa
     1621 cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt
     1681 gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa
     1741 agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata
     1801 ctaacgccgc catccagttt aaacgagctc gaattcatcg atgatatcag atccactagt
     1861 ggcctatgcg gccgcggatc tgccggtctc cctatagtga gtcgtattaa tttcgataag
     1921 ccaggttaac ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1981 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2041 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2101 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2161 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2221 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2281 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2341 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2401 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2461 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2521 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2581 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     2641 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2701 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2761 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2821 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2881 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2941 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3001 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3061 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3121 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3181 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3241 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3301 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3361 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3421 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3481 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3541 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3601 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3661 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3721 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3781 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3841 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3901 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     3961 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     4021 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     4081 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     4141 atcagagcag attgtactga gagtgcacca tatggacata ttgtcgttag aacgcggcta
     4201 caattaatac ataaccttat gtatcataca catacgattt aggtgacact ataga
//
