LOCUS       sequence-454153-        4952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To insert biotin ligase, BASU, at the C-terminus of genes; yeast 
            BioID, HIS3MX marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4952)
  AUTHORS   Pfannenstein J, Tyryshkin M, Gulden ME, Doud EH, Mosley AL, Reese JC
  TITLE     Characterization of BioID tagging systems in budding yeast and 
            exploring the interactome of the Ccr4-Not complex.
  JOURNAL   G3 (Bethesda). 2024 Sep 13:jkae221. doi: 10.1093/g3journal/jkae221.
  PUBMED    39271111
REFERENCE   2  (bases 1 to 4952)
  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: "G3 
            (Bethesda). 2024 Sep 13:jkae221. doi: 10.1093/g3journal/jkae221."
COMMENT     Sequence Label: pFa6-BASU-HA3-HIS3MX
FEATURES             Location/Qualifiers
     source          1..4952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             856..945
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      978..1165
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1033..1057)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1240..1583
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1584..2282
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      2286..2483
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2587..2606)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2588..2606)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2693..2710)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2864..3452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2944..2963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3623..4414,4415..4483))
                     /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     4246..4265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4484..4588)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4656..4674
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4712..4734)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4834..4853
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4934..4952
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4934..4951
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 ggcaagctga gcgaaagtga gattagattc ggtttaaaaa ctgaggtcat gggtcaacat
      121 ttgatatacc atgacgtact gtctagcaca caaaagaccg ctcacgagtt agcaaacaat
      181 aacgctcccg aaggtacatt ggtcgttgcg gataaacaaa cggctggtag aggcggaatg
      241 agtagggtct ggcattcaca agaaggaaat ggtgtgtgga tgtccttgat actaaggcca
      301 gacataccgt tacaaaaaac cccgcaacta acccttttgg ctgcggtggc cgtggttcag
      361 gggatcgagg aagccgctgg gatccaaacg gacataaaat ggccgaatga cattctaatc
      421 aacggcaaaa aaacagtcgg gattctgacc gaaatgcaag ccgaagagga cagggtaaga
      481 agtgtgatca ttgggattgg catcaatgta aatcagcaac ctaatgattt cccggacgaa
      541 ttaaaggaca tagccacctc tcttagccaa gccgctggcg aaaaaataga ccgtgctgga
      601 gtgatccagc atatacttct atgctttgag aagaggtaca gagactacat gactcatggt
      661 tttacgccaa taaagctgtt gtgggaatcc tacgctctgg gaataggaac taacatgcgt
      721 gctagaacac tgaatggtac attttacggt aaggccctag ggattgacga cgaaggtgta
      781 cttcttcttg agacaaacga gggcataaaa aagatataca gtgctgacat atctttgagg
      841 ttaattaaca tcttttaccc atacgatgtt cctgactatg cgggctatcc ctatgacgtc
      901 ccggactatg caggatccta tccatatgac gttccagatt acgctgctca gtgctgaggc
      961 gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta
     1021 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct
     1081 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg
     1141 ctcttattga ccacacctct accggcagat ccgctaggga taacagggta atatagatct
     1201 gtttagcttg cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc
     1261 ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt
     1321 tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg
     1381 cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca
     1441 gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg
     1501 ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac
     1561 atcacatccg aacataaaca accatggcag aaccagccca aaaaaagcaa aaacaaactg
     1621 ttcaggagcg caaggcgttt atctcccgta tcactaatga aactaaaatt caaatcgcta
     1681 tttcgctgaa tggtggttat attcaaataa aagattcgat tcttcctgca aagaaggatg
     1741 acgatgtagc ttcccaagct actcagtcac aggtcatcga tattcacaca ggtgttggct
     1801 ttttggatca tatgatccat gcgttggcaa aacactctgg ttggtctctt attgttgaat
     1861 gtattggtga cctgcacatt gacgatcacc atactaccga agattgcggt atcgcattag
     1921 ggcaagcgtt caaagaagca atgggtgctg tccgtggtgt aaaaagattc ggtactgggt
     1981 tcgcaccatt ggatgaggcg ctatcacgtg ccgtagtcga tttatctagt agaccatttg
     2041 ctgtaatcga ccttggattg aagagagaga tgattggtga tttatccact gaaatgattc
     2101 cacacttttt ggaaagtttc gcggaggcgg ccagaattac tttgcatgtt gattgtctga
     2161 gaggtttcaa cgatcaccac agaagtgaga gtgcgttcaa ggctttggct gttgccataa
     2221 gagaagctat ttctagcaat ggcaccaatg acgttccctc aaccaaaggt gttttgatgt
     2281 gaagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt
     2341 atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg
     2401 acatcatctg cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta
     2461 tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta acgccgccat ccagtttaaa
     2521 cgagctcgaa ttcatcgatg atatcagatc cactagtggc ctatgcggcc gcggatctgc
     2581 cggtctccct atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa
     2641 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2701 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2761 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2821 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2881 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2941 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3001 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3061 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3121 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3181 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3241 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3301 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3361 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3421 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3481 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3541 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3601 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3661 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3721 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3781 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3841 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3901 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3961 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4021 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4081 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4141 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4201 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4261 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4321 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4381 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4441 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4501 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4561 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     4621 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     4681 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     4741 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     4801 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     4861 tgcaccatat ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta
     4921 tcatacacat acgatttagg tgacactata ga
//
