LOCUS       sequence-9645-ex        4142 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 4142)
  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 4142)
  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-3xFLAG-HIS3MX6
FEATURES             Location/Qualifiers
     source          1..4142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..144
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      168..355
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(223..247)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        430..773
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             774..1472
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1476..1673
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1777..1796)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1778..1796)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1883..1900)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2054..2642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2134..2153)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2813..3604,3605..3673))
                     /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     3436..3455
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3674..3778)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3846..3864
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3902..3924)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4024..4043
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4124..4142
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4124..4141
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gggggaggcg ggggtggaga ctacaaagac catgacggtg attataaaga tcatgacatc
      121 gactacaagg atgacgatga caagtagggc gcgccacttc taaataagcg aatttcttat
      181 gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa
      241 gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca
      301 ggttgctttc tcaggtatag tatgaggtcg ctcttattga ccacacctct accggcagat
      361 ccgctaggga taacagggta atatagatct gtttagcttg cctcgtcccc gccgggtcac
      421 ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt gcgcagctca
      481 ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt ataatcattt
      541 gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag
      601 acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc
      661 ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct
      721 tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca accatggcag
      781 aaccagccca aaaaaagcaa aaacaaactg ttcaggagcg caaggcgttt atctcccgta
      841 tcactaatga aactaaaatt caaatcgcta tttcgctgaa tggtggttat attcaaataa
      901 aagattcgat tcttcctgca aagaaggatg acgatgtagc ttcccaagct actcagtcac
      961 aggtcatcga tattcacaca ggtgttggct ttttggatca tatgatccat gcgttggcaa
     1021 aacactctgg ttggtctctt attgttgaat gtatcggtga cctgcacatt gacgatcacc
     1081 atactaccga agattgcggt atcgcattag ggcaagcgtt caaagaagca atgggtgctg
     1141 tccgtggtgt aaaaagattc ggtactgggt tcgcaccatt ggatgaggcg ctatcacgtg
     1201 ccgtagtcga tttatctaat agaccatttg ctgtaatcga ccttggattg aagagagaga
     1261 tgattggtga tttatccact gaaatgattc cacacttttt ggaaagtttc gcggaggcgg
     1321 ccagaattac tttgcatgtt gattgtctga gaggtttcaa cgatcaccac agaagtgaga
     1381 gtgcgttcaa ggctttggct gttgccataa gagaagctat ttctagcaat ggcaccaatg
     1441 acgttccctc aaccaaaggt gttttgatgt gaagtactga caataaaaag attcttgttt
     1501 tcaagaactt gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg
     1561 ttagcgtgat ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg
     1621 cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga
     1681 ttcgatacta acgccgccat ccagtttaaa cgagctcgaa ttcatcgatg atatcagatc
     1741 cactagtggc ctatgcggcc gcggatctgc cggtctccct atagtgagtc gtattaattt
     1801 cgataagcca ggttaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1861 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1921 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1981 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2041 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2101 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2161 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2221 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     2281 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2341 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2401 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2461 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     2521 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2581 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2641 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2701 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2761 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2821 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2881 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2941 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3001 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3061 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3121 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3181 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3241 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3301 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3361 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3421 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3481 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3541 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3601 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3661 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3721 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3781 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     3841 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     3901 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     3961 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     4021 atgcggcatc agagcagatt gtactgagag tgcaccatat ggacatattg tcgttagaac
     4081 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata
     4141 ga
//
