LOCUS       sequence-9671-ex        4228 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 4228)
  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 4228)
  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-kanMX6
FEATURES             Location/Qualifiers
     source          1..4228
                     /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..1759
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        403..746
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             747..1556
                     /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(814..833)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1562..1759
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1863..1882)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1864..1882)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1969..1986)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2140..2728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2220..2239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2899..3690,3691..3759))
                     /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     3522..3541
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3760..3864)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3932..3950
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3988..4010)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4110..4129
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4210..4228
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4210..4227
                     /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 ttgcctcgtc 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 gtaaggaaaa gactcacgtt tcgaggccgc
      781 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
      841 ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc
      901 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
      961 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     1021 catggttact caccactgcg atccccggca aaacagcatt ccaggtatta gaagaatatc
     1081 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     1141 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     1201 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     1261 ctgttgaaca agtctggaaa gaaatgcata agcttttgcc attctcaccg gattcagtcg
     1321 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     1381 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     1441 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     1501 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     1561 tactgacaat aaaaagattc ttgttttcaa gaacttgtca tttgtatagt ttttttatat
     1621 tgtagttgtt ctattttaat caaatgttag cgtgatttat attttttttc gcctcgacat
     1681 catctgccca gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg
     1741 aatgctggtc gctatactgc tgtcgattcg atactaacgc cgccatccag tttaaacgag
     1801 ctcgaattca tcgatgatat cagatccact agtggcctat gcggccgcgg atctgccggt
     1861 ctccctatag tgagtcgtat taatttcgat aagccaggtt aacctgcatt aatgaatcgg
     1921 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     1981 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2041 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2101 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2161 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2221 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2281 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     2341 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2401 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2461 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2521 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2581 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2641 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2701 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2761 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2821 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2881 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2941 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3001 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3061 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3121 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3181 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3241 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3301 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3361 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3421 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3481 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3541 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3601 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3661 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3721 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3781 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3841 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     3901 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     3961 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     4021 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     4081 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     4141 ccatatggac atattgtcgt tagaacgcgg ctacaattaa tacataacct tatgtatcat
     4201 acacatacga tttaggtgac actataga
//
