LOCUS       sequence-9655-ex        4207 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 4207)
  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 4207)
  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-His-tag-kanMX6
FEATURES             Location/Qualifiers
     source          1..4207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..96
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      120..307
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(175..199)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            382..1738
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        382..725
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             726..1535
                     /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(793..812)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1541..1738
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1842..1861)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1843..1861)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1948..1965)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2119..2707)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2199..2218)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2878..3669,3670..3738))
                     /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     3501..3520
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3739..3843)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3911..3929
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3967..3989)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4089..4108
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4189..4207
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4189..4206
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaca tcatcaccat caccactagg gcgcgccact tctaaataag
      121 cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat
      181 acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt
      241 tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct
      301 ctaccggcag atccgctagg gataacaggg taatatagat ctgtttagct tgcctcgtcc
      361 ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac
      421 gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat
      481 gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc
      541 tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc
      601 cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt
      661 catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa
      721 caaccatggg taaggaaaag actcacgttt cgaggccgcg attaaattcc aacatggatg
      781 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
      841 atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
      901 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
      961 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     1021 tccccggcaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     1081 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     1141 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     1201 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     1261 aaatgcataa gcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     1321 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     1381 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     1441 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     1501 tgcagtttca tttgatgctc gatgagtttt tctaatcagt actgacaata aaaagattct
     1561 tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc tattttaatc
     1621 aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag atgcgaagtt
     1681 aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg ctatactgct
     1741 gtcgattcga tactaacgcc gccatccagt ttaaacgagc tcgaattcat cgatgatatc
     1801 agatccacta gtggcctatg cggccgcgga tctgccggtc tccctatagt gagtcgtatt
     1861 aatttcgata agccaggtta acctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1921 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1981 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2041 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2101 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2161 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2221 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2281 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     2341 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2401 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2461 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2521 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     2581 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2641 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2701 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2761 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2821 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2881 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2941 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3001 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3061 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3121 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3181 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3241 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3301 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3361 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3421 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3481 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3541 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3601 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3661 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3721 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3781 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3841 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     3901 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     3961 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     4021 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     4081 taactatgcg gcatcagagc agattgtact gagagtgcac catatggaca tattgtcgtt
     4141 agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca
     4201 ctataga
//
