LOCUS       sequence-244942-        5227 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For C-terminal tagging with TurboID in yeast including 
            geneticin-resistant marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5227)
  AUTHORS   Larochelle M, Bergeron D, Arcand B, Bachand F
  TITLE     Proximity-dependent biotinylation by TurboID to identify 
            protein-protein interaction networks in yeast.
  JOURNAL   J Cell Sci. 2019 May 7. pii: jcs.232249. doi: 10.1242/jcs.232249.
  PUBMED    31064814
REFERENCE   2  (bases 1 to 5227)
  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: "J Cell Sci.
            2019 May 7. pii: jcs.232249. doi: 10.1242/jcs.232249."
COMMENT     Sequence Label: pFA6a-TurboID-3MYC-kanMX6 (pFB1420)
FEATURES             Location/Qualifiers
     source          1..5227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1027..1116
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     terminator      1140..1327
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1195..1219)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1402..2758
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1402..1745
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1746..2555
                     /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(1813..1832)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2561..2758
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2862..2881)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2863..2881)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2968..2985)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3139..3727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3219..3238)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3898..4689,4690..4758))
                     /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     4521..4540
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4759..4863)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4931..4949
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4987..5009)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5109..5128
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5209..5227
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5209..5226
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 atctttaaag acaatacagt gccgctcaaa ttgatagccc tccttgcaaa tggagagttt
      121 cacagcggtg aacaactggg cgagactctg ggtatgtcac gcgcagctat aaataagcat
      181 atacagacac tcagggattg gggggttgac gtatttacag tgccaggcaa gggctattct
      241 ctcccagagc ccataccact tttgaatgct aaacagatac ttggtcaatt ggatggaggt
      301 agcgttgcgg tgctcccggt tgttgattcc acaaatcagt atctgctgga taggattgga
      361 gaactcaaaa gcggggatgc ctgtatcgcg gaataccaac aagctggtcg aggctcacga
      421 ggcagaaaat ggttctcccc gttcggggca aatctgtacc ttagcatgtt ttggcgcttg
      481 aaaagaggac ccgcggcgat aggactcggc cccgttattg gaattgttat ggccgaagct
      541 cttcgaaaat tgggtgctga caaggttcgg gtgaagtggc ctaacgattt gtatttgcag
      601 gaccggaaac tcgcaggaat tttggtcgag ctggcaggta ttactggtga cgcggctcaa
      661 attgttatag gtgccggcat caacgtcgct atgagacggg tcgaagagtc cgttgtaaat
      721 caagggtgga ttacactcca ggaggcaggc ataaatctcg accgaaatac gttggctgct
      781 accctgattc gagagcttag agctgcactc gaattgtttg aacaagaagg tctcgcacca
      841 tatcttccga gatgggagaa actcgataat ttcattaacc ggcctgtcaa gttgatcatt
      901 ggagataaag agattttcgg gatttccaga ggtattgaca aacaaggcgc tctcctcctt
      961 gagcaagatg gggtgatcaa gccttggatg gggggcgaga tctccctgag gtcagcagag
     1021 aaagacgagc agaaactcat ctctgaagaa gatctggaac aaaagttgat ttcagaagaa
     1081 gatctggaac agaagctcat ctctgaggaa gatctgtgag gcgcgccact tctaaataag
     1141 cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat
     1201 acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt
     1261 tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct
     1321 ctaccggcag atccgctagg gataacaggg taatatagat ctgtttagct tgcctcgtcc
     1381 ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac
     1441 gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat
     1501 gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc
     1561 tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc
     1621 cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt
     1681 catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa
     1741 caaccatggg taaggaaaag actcacgttt cgaggccgcg attaaattcc aacatggatg
     1801 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     1861 atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     1921 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     1981 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     2041 tccccggcaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     2101 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     2161 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     2221 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     2281 aaatgcataa gcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     2341 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     2401 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     2461 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     2521 tgcagtttca tttgatgctc gatgagtttt tctaatcagt actgacaata aaaagattct
     2581 tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc tattttaatc
     2641 aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag atgcgaagtt
     2701 aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg ctatactgct
     2761 gtcgattcga tactaacgcc gccatccagt ttaaacgagc tcgaattcat cgatgatatc
     2821 agatccacta gtggcctatg cggccgcgga tctgccggtc tccctatagt gagtcgtatt
     2881 aatttcgata agccaggtta acctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2941 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3001 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3061 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3121 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3181 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3241 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3301 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3361 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3421 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3481 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3541 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3601 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3661 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3721 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3781 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3841 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3901 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3961 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4021 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4081 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4141 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4201 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4261 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4321 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4381 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4441 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4501 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4561 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4621 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4681 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4741 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4801 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4861 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4921 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     4981 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5041 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5101 taactatgcg gcatcagagc agattgtact gagagtgcac catatggaca tattgtcgtt
     5161 agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca
     5221 ctataga
//
