LOCUS       sequence-454159-        4976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To insert biotin ligase, miniTurboID, 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 4976)
  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 4976)
  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-mTID-HA3-HIS3MX6
FEATURES             Location/Qualifiers
     source          1..4976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             880..969
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      1002..1189
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1057..1081)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1264..1607
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1608..2306
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      2310..2507
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2611..2630)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2612..2630)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2717..2734)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2888..3476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2968..2987)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3647..4438,4439..4507))
                     /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     4270..4289
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4508..4612)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4680..4698
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4736..4758)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4858..4877
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4958..4976
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4958..4975
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gcaagagacc cgcccgtagc aactatgatt cctttactga atgcgaaaca gatattgggt
      121 caattagatg ggggttccgt agctgtcctt ccggtggtgg atagcacgaa ccaatatcta
      181 cttgatagga tcggggaatt gaagtcaggt gatgcttgca tcgcagagta ccaacaggct
      241 ggtagaggga gcagaggcag gaagtggttt tcaccatttg gagcaaatct gtacttgagc
      301 atgttctgga gacttaagag aggtccagcg gcgatagggc tgggacccgt tatcggaatt
      361 gtgatggcag aagccctgcg taaactaggt gctgataaag tacgtgtcaa atggccaaac
      421 gatttatatc tacaagacag aaagttggcg gggattttgg tggaacttgc cggcataacc
      481 ggggacgccg cgcagatagt cattggtgct ggaatcaatg tggcgatgag acgtgttgag
      541 gagtcagttg taaatcaggg ttggataact cttcaggagg ccgggataaa cttagacagg
      601 aatactctag ctgccatgct tatacgtgag ctacgtgccg ccctggagct tttcgagcaa
      661 gaaggtctag ctccttacct tagccgttgg gagaagttgg ataacttcat caatagacca
      721 gttaagttaa tcatagggga caaagagata ttcgggatca gtagaggtat agataagcag
      781 ggagcgctac tactggagca ggatggggtt atcaaaccgt ggatgggcgg ggagatatcc
      841 ctgaggtctg ccgagaaggc tagcttaatt aacatctttt acccatacga tgttcctgac
      901 tatgcgggct atccctatga cgtcccggac tatgcaggat cctatccata tgacgttcca
      961 gattacgctg ctcagtgctg aggcgcgcca cttctaaata agcgaatttc ttatgattta
     1021 tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact
     1081 cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc
     1141 tttctcaggt atagtatgag gtcgctctta ttgaccacac ctctaccggc agatccgcta
     1201 gggataacag ggtaatatag atctgtttag cttgcctcgt ccccgccggg tcacccggcc
     1261 agcgacatgg aggcccagaa taccctcctt gacagtcttg acgtgcgcag ctcaggggca
     1321 tgatgtgact gtcgcccgta catttagccc atacatcccc atgtataatc atttgcatcc
     1381 atacattttg atggccgcac ggcgcgaagc aaaaattacg gctcctcgct gcagacctgc
     1441 gagcagggaa acgctcccct cacagacgcg ttgaattgtc cccacgccgc gcccctgtag
     1501 agaaatataa aaggttagga tttgccactg aggttcttct ttcatatact tccttttaaa
     1561 atcttgctag gatacagttc tcacatcaca tccgaacata aacaaccatg gcagaaccag
     1621 cccaaaaaaa gcaaaaacaa actgttcagg agcgcaaggc gtttatctcc cgtatcacta
     1681 atgaaactaa aattcaaatc gctatttcgc tgaatggtgg ttatattcaa ataaaagatt
     1741 cgattcttcc tgcaaagaag gatgacgatg tagcttccca agctactcag tcacaggtca
     1801 tcgatattca cacaggtgtt ggctttttgg atcatatgat ccatgcgttg gcaaaacact
     1861 ctggttggtc tcttattgtt gaatgtattg gtgacctgca cattgacgat caccatacta
     1921 ccgaagattg cggtatcgca ttagggcaag cgttcaaaga agcaatgggt gctgtccgtg
     1981 gtgtaaaaag attcggtact gggttcgcac cattggatga ggcgctatca cgtgccgtag
     2041 tcgatttatc tagtagacca tttgctgtaa tcgaccttgg attgaagaga gagatgattg
     2101 gtgatttatc cactgaaatg attccacact ttttggaaag tttcgcggag gcggccagaa
     2161 ttactttgca tgttgattgt ctgagaggtt tcaacgatca ccacagaagt gagagtgcgt
     2221 tcaaggcttt ggctgttgcc ataagagaag ctatttctag caatggcacc aatgacgttc
     2281 cctcaaccaa aggtgttttg atgtgaagta ctgacaataa aaagattctt gttttcaaga
     2341 acttgtcatt tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg
     2401 tgatttatat tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga
     2461 aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat
     2521 actaacgccg ccatccagtt taaacgagct cgaattcatc gatgatatca gatccactag
     2581 tggcctatgc ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa
     2641 gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2701 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2761 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2821 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2881 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2941 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3001 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3061 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3121 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3181 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3241 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3301 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3361 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3421 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3481 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3541 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3601 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3661 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3721 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3781 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3841 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3901 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3961 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4021 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4081 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4141 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4201 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4261 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4321 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4381 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4441 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4501 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4561 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4621 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4681 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     4741 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     4801 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     4861 catcagagca gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct
     4921 acaattaata cataacctta tgtatcatac acatacgatt taggtgacac tataga
//
