LOCUS       sequence-9652-ex        4222 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 4222)
  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 4222)
  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-T7-kanMX6
FEATURES             Location/Qualifiers
     source          1..4222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..111
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     terminator      135..322
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(190..214)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            397..1753
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        397..740
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             741..1550
                     /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(808..827)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1556..1753
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1857..1876)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1858..1876)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1963..1980)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2134..2722)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2214..2233)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2893..3684,3685..3753))
                     /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     3516..3535
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3754..3858)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3926..3944
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3982..4004)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4104..4123
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4204..4222
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4204..4221
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaat ggctagcatg actggtggac agcaaatggg ttagggcgcg
      121 ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
      181 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
      241 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc
      301 ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata tagatctgtt
      361 tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
      421 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
      481 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
      541 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
      601 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
      661 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
      721 acatccgaac ataaacaacc atgggtaagg aaaagactca cgtttcgagg ccgcgattaa
      781 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
      841 caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac
      901 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
      961 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
     1021 tactcaccac tgcgatcccc ggcaaaacag cattccaggt attagaagaa tatcctgatt
     1081 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
     1141 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
     1201 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     1261 aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca gtcgtcactc
     1321 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     1381 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     1441 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     1501 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagtactga
     1561 caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt atattgtagt
     1621 tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg acatcatctg
     1681 cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct
     1741 ggtcgctata ctgctgtcga ttcgatacta acgccgccat ccagtttaaa cgagctcgaa
     1801 ttcatcgatg atatcagatc cactagtggc ctatgcggcc gcggatctgc cggtctccct
     1861 atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa tcggccaacg
     1921 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     1981 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2041 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2101 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2161 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2221 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2281 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     2341 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2401 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2461 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2521 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     2581 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2641 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2701 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2761 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2821 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2881 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2941 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3001 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3061 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3121 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3181 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3241 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3301 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3361 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3421 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3481 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3541 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3601 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3661 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3721 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3781 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3841 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     3901 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     3961 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     4021 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     4081 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     4141 ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat
     4201 acgatttagg tgacactata ga
//
