LOCUS       sequence-454141-        5507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To integrate TID-FKBP-NLS fusion at yeast promoter; yeast BioID with
            HISMX6 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5507)
  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 5507)
  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-NLS-3HA-HISMX6
FEATURES             Location/Qualifiers
     source          1..5507
                     /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             1396..1416
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1417..1506
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      1533..1720
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1588..1612)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1795..2138
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2139..2837
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      2841..3038
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3142..3161)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3143..3161)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3248..3265)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3419..4007)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3499..3518)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4178..4969,4970..5038))
                     /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     4801..4820
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5039..5143)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5211..5229
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5267..5289)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5389..5408
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5489..5507
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5489..5506
                     /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 ctccaccaaa aaagaaaaga aaagtttacc catacgatgt tcctgactat
     1441 gcgggctatc cgtatgacgt cccggactat gcaggatctt atccatatga cgttccagat
     1501 tacgctgctt gaggcgcgcc acttctaaat aagcgaattt cttatgattt atgattttta
     1561 ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt
     1621 taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg
     1681 tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct agggataaca
     1741 gggtaatata gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg
     1801 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac
     1861 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt
     1921 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga
     1981 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata
     2041 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta
     2101 ggatacagtt ctcacatcac atccgaacat aaacaaccat ggcagaacca gcccaaaaaa
     2161 agcaaaaaca aactgttcag gagcgcaagg cgtttatctc ccgtatcact aatgaaacta
     2221 aaattcaaat cgctatttcg ctgaatggtg gttatattca aataaaagat tcgattcttc
     2281 ctgcaaagaa ggatgacgat gtagcttccc aagctactca gtcacaggtc atcgatattc
     2341 acacaggtgt tggctttttg gatcatatga tccatgcgtt ggcaaaacac tctggttggt
     2401 ctcttattgt tgaatgtatt ggtgacctgc acattgacga tcaccatact accgaagatt
     2461 gcggtatcgc attagggcaa gcgttcaaag aagcaatggg tgctgtccgt ggtgtaaaaa
     2521 gattcggtac tgggttcgca ccattggatg aggcgctatc acgtgccgta gtcgatttat
     2581 ctagtagacc atttgctgta atcgaccttg gattgaagag agagatgatt ggtgatttat
     2641 ccactgaaat gattccacac tttttggaaa gtttcgcgga ggcggccaga attactttgc
     2701 atgttgattg tctgagaggt ttcaacgatc accacagaag tgagagtgcg ttcaaggctt
     2761 tggctgttgc cataagagaa gctatttcta gcaatggcac caatgacgtt ccctcaacca
     2821 aaggtgtttt gatgtgaagt actgacaata aaaagattct tgttttcaag aacttgtcat
     2881 ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata
     2941 ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat
     3001 catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc
     3061 gccatccagt ttaaacgagc tcgaattcat cgatgatatc agatccacta gtggcctatg
     3121 cggccgcgga tctgccggtc tccctatagt gagtcgtatt aatttcgata agccaggtta
     3181 acctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3241 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3301 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3361 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3421 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3481 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3541 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3601 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3661 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3721 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3781 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3841 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3901 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3961 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4021 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4081 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4141 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4201 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4261 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4321 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4381 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4441 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4501 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4561 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4621 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4681 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4741 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4801 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4861 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4921 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4981 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5041 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5101 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5161 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5221 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     5281 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     5341 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     5401 agattgtact gagagtgcac catatggaca tattgtcgtt agaacgcggc tacaattaat
     5461 acataacctt atgtatcata cacatacgat ttaggtgaca ctataga
//
