LOCUS       sequence-454151-        5162 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To insert biotin ligase, TurboID, at the C-terminus of genes; yeast 
            BioID, HIS3MX6 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5162)
  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 5162)
  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: pFA6a-TID-HA3-HIS3MX6
FEATURES             Location/Qualifiers
     source          1..5162
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1066..1155
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      1188..1375
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1243..1267)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1450..1793
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1794..2492
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      2496..2693
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2797..2816)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2798..2816)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2903..2920)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3074..3662)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3154..3173)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3833..4624,4625..4693))
                     /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     4456..4475
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4694..4798)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4866..4884
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4922..4944)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5044..5063
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5144..5162
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5144..5161
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gcaagagatc cgcctgtggc cacgatgaaa gataacacgg ttcccttaaa attgatagcc
      121 cttcttgcca atggtgagtt tcattccggg gaacaacttg gcgaaactct gggaatgagc
      181 agggcagcga ttaacaagca tatccaaacc ttgagagact ggggagtaga tgtgttcacc
      241 gtgcctggta aaggttattc tttgccggaa cctatccctc tattaaatgc gaaacagata
      301 ctaggacaac tggacggtgg tagtgttgcc gtattgccgg ttgttgactc aaccaatcaa
      361 tacttgctgg acagaattgg cgaacttaaa agcggtgatg cgtgtattgc agaatatcag
      421 caagcaggga gaggctcaag aggtaggaaa tggttttcac cttttggcgc caatctttac
      481 ttaagtatgt tctggcgttt gaagagaggg cccgcggcta ttggtttagg tcccgttatc
      541 gggatagtta tggctgaagc gctacgtaaa cttggggccg acaaggtcag agtgaagtgg
      601 cctaatgatc tatacttgca agatagaaag ctagcgggta tcctggtgga gctagcggga
      661 atcacagggg acgctgccca aatagtaatc ggtgctggta taaatgtcgc aatgcgtaga
      721 gtcgaggaat cagtggtgaa ccagggctgg atcacgttgc aagaggcggg cataaacctg
      781 gaccgtaaca cccttgcagc gaccctaata agagaactac gtgctgcatt ggagcttttt
      841 gagcaggagg gcctggctcc gtacctaccc aggtgggaga aattagacaa tttcataaac
      901 cgtccagtca aactaataat tggagataag gagattttcg gtatttctcg tgggatagac
      961 aagcagggag cgttgttatt agaacaggat ggtgtcatta aaccttggat gggcggtgaa
     1021 attagtcttc gtagtgccga aaaggctagc ttaattaaca tcttttaccc atacgatgtt
     1081 cctgactatg cgggctatcc ctatgacgtc ccggactatg caggatccta tccatatgac
     1141 gttccagatt acgctgctca gtgctgaggc gcgccacttc taaataagcg aatttcttat
     1201 gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa
     1261 gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca
     1321 ggttgctttc tcaggtatag tatgaggtcg ctcttattga ccacacctct accggcagat
     1381 ccgctaggga taacagggta atatagatct gtttagcttg cctcgtcccc gccgggtcac
     1441 ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt gcgcagctca
     1501 ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt ataatcattt
     1561 gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag
     1621 acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc
     1681 ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct
     1741 tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca accatggcag
     1801 aaccagccca aaaaaagcaa aaacaaactg ttcaggagcg caaggcgttt atctcccgta
     1861 tcactaatga aactaaaatt caaatcgcta tttcgctgaa tggtggttat attcaaataa
     1921 aagattcgat tcttcctgca aagaaggatg acgatgtagc ttcccaagct actcagtcac
     1981 aggtcatcga tattcacaca ggtgttggct ttttggatca tatgatccat gcgttggcaa
     2041 aacactctgg ttggtctctt attgttgaat gtattggtga cctgcacatt gacgatcacc
     2101 atactaccga agattgcggt atcgcattag ggcaagcgtt caaagaagca atgggtgctg
     2161 tccgtggtgt aaaaagattc ggtactgggt tcgcaccatt ggatgaggcg ctatcacgtg
     2221 ccgtagtcga tttatctagt agaccatttg ctgtaatcga ccttggattg aagagagaga
     2281 tgattggtga tttatccact gaaatgattc cacacttttt ggaaagtttc gcggaggcgg
     2341 ccagaattac tttgcatgtt gattgtctga gaggtttcaa cgatcaccac agaagtgaga
     2401 gtgcgttcaa ggctttggct gttgccataa gagaagctat ttctagcaat ggcaccaatg
     2461 acgttccctc aaccaaaggt gttttgatgt gaagtactga caataaaaag attcttgttt
     2521 tcaagaactt gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg
     2581 ttagcgtgat ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg
     2641 cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga
     2701 ttcgatacta acgccgccat ccagtttaaa cgagctcgaa ttcatcgatg atatcagatc
     2761 cactagtggc ctatgcggcc gcggatctgc cggtctccct atagtgagtc gtattaattt
     2821 cgataagcca ggttaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2881 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2941 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3001 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3061 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3121 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3181 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3241 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3301 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3361 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3421 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3481 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3541 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3601 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3661 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3721 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3781 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3841 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3901 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3961 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4021 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4081 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4141 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4201 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4261 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4321 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4381 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4441 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4501 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4561 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4621 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4681 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4741 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4801 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     4861 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     4921 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     4981 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     5041 atgcggcatc agagcagatt gtactgagag tgcaccatat ggacatattg tcgttagaac
     5101 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata
     5161 ga
//
