LOCUS       sequence-9643-ex        4213 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 4213)
  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 4213)
  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-FLAG-kanMX6
FEATURES             Location/Qualifiers
     source          1..4213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..102
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      126..313
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(181..205)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            388..1744
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        388..731
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             732..1541
                     /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(799..818)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1547..1744
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1848..1867)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1849..1867)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1954..1971)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2125..2713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2205..2224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2884..3675,3676..3744))
                     /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     3507..3526
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3745..3849)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3917..3935
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3973..3995)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4095..4114
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4195..4213
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4195..4212
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaga ttacaaggac gacgatgaca agtagggcgc gccacttcta
      121 aataagcgaa tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa
      181 gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta ttcttgagta
      241 actctttcct gtaggtcagg ttgctttctc aggtatagta tgaggtcgct cttattgacc
      301 acacctctac cggcagatcc gctagggata acagggtaat atagatctgt ttagcttgcc
      361 tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct ccttgacagt
      421 cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta gcccatacat
      481 ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat
      541 tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat
      601 tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc actgaggttc
      661 ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat cacatccgaa
      721 cataaacaac catgggtaag gaaaagactc acgtttcgag gccgcgatta aattccaaca
      781 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
      841 caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
      901 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
      961 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     1021 ctgcgatccc cggcaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     1081 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     1141 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     1201 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     1261 ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt
     1321 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     1381 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     1441 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     1501 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagtactg acaataaaaa
     1561 gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt
     1621 ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc
     1681 gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat
     1741 actgctgtcg attcgatact aacgccgcca tccagtttaa acgagctcga attcatcgat
     1801 gatatcagat ccactagtgg cctatgcggc cgcggatctg ccggtctccc tatagtgagt
     1861 cgtattaatt tcgataagcc aggttaacct gcattaatga atcggccaac gcgcggggag
     1921 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1981 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2041 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2101 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2161 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2221 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2281 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     2341 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2401 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2461 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2521 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     2581 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2641 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2701 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2761 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2821 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2881 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2941 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3001 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3061 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3121 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3181 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3241 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3301 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3361 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3421 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3481 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3541 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3601 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3661 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3721 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3781 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3841 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     3901 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     3961 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     4021 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     4081 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tggacatatt
     4141 gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag
     4201 gtgacactat aga
//
