LOCUS       sequence-451860-        5596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To integrate TID-FKBP fusion at yeast promoter; yeast BioID with 
            KanMX6 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5596)
  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 5596)
  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-TID-FKBP-3HA-KanMX6
FEATURES             Location/Qualifiers
     source          1..5596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1069..1389
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1393..1482
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      1509..1696
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1564..1588)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1771..3127
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1771..2114
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2115..2924
                     /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(2182..2201)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2930..3127
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3231..3250)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3232..3250)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3337..3354)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3508..4096)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3588..3607)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4267..5058,5059..5127))
                     /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     4890..4909
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5128..5232)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5300..5318
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5356..5378)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5478..5497
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5578..5596
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5578..5595
                     /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 ggttctggtg gtgctagcgg agtgcaggtg
     1081 gaaaccatct ccccaggaga cgggcgcacc ttccccaagc gcggccagac ctgcgtggtg
     1141 cactacaccg ggatgcttga agatggaaag aaatttgatt cctcccggga cagaaacaag
     1201 ccctttaagt ttatgctagg caagcaggag gtgatccgag gctgggaaga aggggttgcc
     1261 cagatgagtg tgggtcagag agccaaactg actatatctc cagattatgc ctatggtgcc
     1321 actgggcacc caggcatcat cccaccacat gccactctcg tcttcgatgt ggagcttcta
     1381 aaactggaaa cttacccata cgatgttcct gactatgcgg gctatccgta tgacgtcccg
     1441 gactatgcag gatcttatcc atatgacgtt ccagattacg ctgcttgagg cgcgccactt
     1501 ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa
     1561 taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga
     1621 gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc gctcttattg
     1681 accacacctc taccggcaga tccgctaggg ataacagggt aatatagatc tgtttagctt
     1741 gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac
     1801 agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata
     1861 catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa
     1921 aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg
     1981 aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg
     2041 ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc
     2101 gaacataaac aaccatgggt aaggaaaaga ctcacgtttc gaggccgcga ttaaattcca
     2161 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     2221 cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     2281 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     2341 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     2401 ccactgcgat ccccggcaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     2461 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     2521 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     2581 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     2641 tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     2701 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     2761 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     2821 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     2881 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagta ctgacaataa
     2941 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
     3001 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
     3061 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc
     3121 tatactgctg tcgattcgat actaacgccg ccatccagtt taaacgagct cgaattcatc
     3181 gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg
     3241 agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg
     3301 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3361 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3421 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3481 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3541 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3601 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3661 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3721 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3781 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3841 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3901 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3961 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4021 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4081 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4141 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4201 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4261 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4321 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4381 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4441 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4501 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4561 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4621 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4681 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4741 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4801 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4861 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4921 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4981 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5041 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5101 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5161 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5221 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     5281 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     5341 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5401 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     5461 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat
     5521 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
     5581 taggtgacac tataga
//
