LOCUS       sequence-49547-e        4276 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 4276)
  AUTHORS   Bahler J, Wu JQ, Longtine MS, Shah NG, McKenzie A 3rd, Steever AB, 
            Wach A, Philippsen P, Pringle JR
  TITLE     Heterologous modules for efficient and versatile PCR-based gene 
            targeting in Schizosaccharomyces pombe.
  JOURNAL   Yeast. 1998 Jul;14(10):943-51.
  PUBMED    9717240
REFERENCE   2  (bases 1 to 4276)
  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."; 
            date: "1998-07"; volume: "14(10)"; pages: "943-51"
COMMENT     Sequence Label: pFA6a-3HA-kanMX6
FEATURES             Location/Qualifiers
     source          1..4276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             67..156
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      189..376
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(244..268)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            451..1807
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        451..794
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             795..1604
                     /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(862..881)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1610..1807
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1911..1930)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1912..1930)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2017..2034)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2188..2776)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2268..2287)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2947..3738,3739..3807))
                     /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     3570..3589
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3808..3912)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3980..3998
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4036..4058)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4158..4177
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4258..4276
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4258..4275
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 atcttttacc catacgatgt tcctgactat gcgggctatc cgtatgacgt cccggactat
      121 gcaggatcct atccatatga cgttccagat tacgctgctc agtgctgagg cgcgccactt
      181 ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa
      241 taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga
      301 gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc gctcttattg
      361 accacacctc taccggcaga tccgctaggg ataacagggt aatatagatc tgtttagctt
      421 gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac
      481 agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata
      541 catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa
      601 aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg
      661 aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg
      721 ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc
      781 gaacataaac aaccatgggt aaggaaaaga ctcacgtttc gaggccgcga ttaaattcca
      841 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
      901 cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
      961 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     1021 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     1081 ccactgcgat ccccggcaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     1141 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     1201 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     1261 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     1321 tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     1381 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     1441 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     1501 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     1561 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagta ctgacaataa
     1621 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
     1681 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
     1741 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc
     1801 tatactgctg tcgattcgat actaacgccg ccatccagtt taaacgagct cgaattcatc
     1861 gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg
     1921 agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg
     1981 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2041 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2101 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2161 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2221 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2281 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2341 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     2401 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2461 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2521 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2581 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     2641 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2701 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2761 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2821 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2881 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2941 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3001 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3061 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3121 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3181 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3241 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3301 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3361 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3421 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3481 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3541 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3601 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3661 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3721 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3781 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     3841 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3901 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     3961 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4021 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4081 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4141 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat
     4201 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
     4261 taggtgacac tataga
//
