LOCUS       sequence-9670-ex        4450 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 4450)
  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 4450)
  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-V5-hphMX4
FEATURES             Location/Qualifiers
     source          1..4450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             76..117
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     terminator      141..328
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(196..220)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            403..1978
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        403..746
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             747..1775
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             747..1775
                     /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      1781..1978
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2085..2104)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2086..2104)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2191..2208)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2362..2950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2442..2461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3121..3912,3913..3981))
                     /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     3744..3763
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3982..4086)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4154..4172
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4210..4232)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4332..4351
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4432..4450
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4432..4449
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaaa gcctatccct aaccctctcc tcggtctcga ttctacgtag
      121 ggcgcgccac ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag
      181 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
      241 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg
      301 tcgctcttat tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga
      361 tctgtttagc ttgccttgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat
      421 accctccttg acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac
      481 atttagccca tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg
      541 gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc
      601 acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat
      661 ttgccactga ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct
      721 cacatcacat ccgaacataa acaaccatgg gtaaaaagcc tgaactcacc gcgacgtctg
      781 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
      841 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
      901 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg
      961 cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg acctattgca
     1021 tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
     1081 ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga
     1141 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
     1201 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     1261 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     1321 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
     1381 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
     1441 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
     1501 ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc
     1561 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
     1621 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
     1681 gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
     1741 gacgccccag cactcgtccg agggcaaagg aataatcagt actgacaata aaaagattct
     1801 tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc tattttaatc
     1861 aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag atgcgaagtt
     1921 aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg ctatactgct
     1981 gtcgattcga tactaacgcc gccatccagt gtcgaaaacg agctcgaatt catcgatgat
     2041 atcagatcca ctagtggcct atgcggccgc ggatctgccg gtctccctat agtgagtcgt
     2101 attaatttcg ataagccagg ttaacctgca ttaatgaatc ggccaacgcg cggggagagg
     2161 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2221 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2281 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2341 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2401 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2461 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2521 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     2581 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2641 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2701 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2761 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     2821 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2881 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2941 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3001 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3061 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3121 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3181 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3241 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3301 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3361 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3421 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3481 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3541 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3601 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3661 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3721 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3781 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3841 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3901 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3961 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4021 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4081 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4141 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     4201 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     4261 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     4321 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgg acatattgtc
     4381 gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg
     4441 acactataga
//
