LOCUS       sequence-454142-        5483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To integrate TID-FKBP fusion at yeast promoter; yeast BioID with 
            HIS3MX6 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5483)
  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 5483)
  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-HIS3MX6
FEATURES             Location/Qualifiers
     source          1..5483
                     /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"
     promoter        1771..2114
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2115..2813
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      2817..3014
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3118..3137)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3119..3137)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3224..3241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3395..3983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3475..3494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4154..4945,4946..5014))
                     /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     4777..4796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5015..5119)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5187..5205
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5243..5265)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5365..5384
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5465..5483
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5465..5482
                     /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 aaccatggca gaaccagccc aaaaaaagca aaaacaaact gttcaggagc
     2161 gcaaggcgtt tatctcccgt atcactaatg aaactaaaat tcaaatcgct atttcgctga
     2221 atggtggtta tattcaaata aaagattcga ttcttcctgc aaagaaggat gacgatgtag
     2281 cttcccaagc tactcagtca caggtcatcg atattcacac aggtgttggc tttttggatc
     2341 atatgatcca tgcgttggca aaacactctg gttggtctct tattgttgaa tgtattggtg
     2401 acctgcacat tgacgatcac catactaccg aagattgcgg tatcgcatta gggcaagcgt
     2461 tcaaagaagc aatgggtgct gtccgtggtg taaaaagatt cggtactggg ttcgcaccat
     2521 tggatgaggc gctatcacgt gccgtagtcg atttatctag tagaccattt gctgtaatcg
     2581 accttggatt gaagagagag atgattggtg atttatccac tgaaatgatt ccacactttt
     2641 tggaaagttt cgcggaggcg gccagaatta ctttgcatgt tgattgtctg agaggtttca
     2701 acgatcacca cagaagtgag agtgcgttca aggctttggc tgttgccata agagaagcta
     2761 tttctagcaa tggcaccaat gacgttccct caaccaaagg tgttttgatg tgaagtactg
     2821 acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag
     2881 ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct
     2941 gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc
     3001 tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtttaa acgagctcga
     3061 attcatcgat gatatcagat ccactagtgg cctatgcggc cgcggatctg ccggtctccc
     3121 tatagtgagt cgtattaatt tcgataagcc aggttaacct gcattaatga atcggccaac
     3181 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3241 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3301 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3361 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3421 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3481 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3541 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3601 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3661 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3721 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3781 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3841 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3901 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3961 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4021 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4081 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4141 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4201 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4261 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4321 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4381 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4441 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4501 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4561 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4621 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4681 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4741 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4801 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4861 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4921 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4981 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5041 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5101 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     5161 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     5221 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     5281 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     5341 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     5401 tggacatatt gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca
     5461 tacgatttag gtgacactat aga
//
