LOCUS       sequence-9659-ex        4456 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 4456)
  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 4456)
  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-S-tag-hphMX4
FEATURES             Location/Qualifiers
     source          1..4456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..123
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      147..334
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(202..226)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            409..1984
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        409..752
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             753..1781
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             753..1781
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      1787..1984
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2091..2110)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2092..2110)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2197..2214)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2368..2956)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2448..2467)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3127..3918,3919..3987))
                     /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     3750..3769
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3988..4092)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4160..4178
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4216..4238)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4338..4357
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4438..4456
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4438..4455
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaaa agaaaccgct gctgctaaat tcgaacgcca gcacatggac
      121 agctagggcg cgccacttct aaataagcga atttcttatg atttatgatt tttattatta
      181 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
      241 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt
      301 atgaggtcgc tcttattgac cacacctcta ccggcagatc cgctagggat aacagggtaa
      361 tatagatctg tttagcttgc cttgtccccg ccgggtcacc cggccagcga catggaggcc
      421 cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc
      481 ccgtacattt agcccataca tccccatgta taatcatttg catccataca ttttgatggc
      541 cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca gggaaacgct
      601 cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt
      661 taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac
      721 agttctcaca tcacatccga acataaacaa ccatgggtaa aaagcctgaa ctcaccgcga
      781 cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct
      841 cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc
      901 gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat
      961 cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct
     1021 attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc
     1081 ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc
     1141 agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg
     1201 atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca
     1261 ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc
     1321 ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg
     1381 gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg
     1441 tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact
     1501 tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat atgctccgca
     1561 ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg
     1621 cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa
     1681 tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg
     1741 gaaaccgacg ccccagcact cgtccgaggg caaaggaata atcagtactg acaataaaaa
     1801 gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt
     1861 ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc
     1921 gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat
     1981 actgctgtcg attcgatact aacgccgcca tccagtgtcg aaaacgagct cgaattcatc
     2041 gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg
     2101 agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg
     2161 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2221 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2281 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2341 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2401 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2461 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2521 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     2581 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2641 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2701 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2761 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     2821 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2881 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2941 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3001 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3061 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3121 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3181 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3241 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3301 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3361 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3421 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3481 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3541 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3601 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3661 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3721 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3781 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3841 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3901 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3961 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4021 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4081 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4141 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4201 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4261 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4321 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat
     4381 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
     4441 taggtgacac tataga
//
