LOCUS       sequence-178255-        8585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Split-BioID plasmid: Expresses C-terminally tagged CBir[257-321]-FRB
            and C-terminally tagged NBir[2-256]-FKBP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8585)
  AUTHORS   Schopp IM, Amaya Ramirez CC, Debeljak J, Kreibich E, Skribbe M, Wild
            K, Bethune J
  TITLE     Split-BioID a conditional proteomics approach to monitor the 
            composition of spatiotemporally defined protein complexes.
  JOURNAL   Nat Commun. 2017 Jun 6;8:15690. doi: 10.1038/ncomms15690.
  PUBMED    28585547
REFERENCE   2  (bases 1 to 8585)
  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: "Nat 
            Commun."; date: "2017-06-6"; volume: "8:15690. doi"; pages: " 
            10.1038/ncomms15690"
COMMENT     Sequence Label: pSF3-Flag-FRB-Cbir_FKBP-Myc-Nbir
FEATURES             Location/Qualifiers
     source          1..8585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             402..995
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    1058..1415
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     primer_bind     1826..1848
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1864..1880
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    1933..1966
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     polyA_signal    2074..2208
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2111..2130)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3163..3192)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(join(3253..3573,3574..3576))
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     promoter        complement(3680..3717)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    join(3734..3752,3753..3775,3776..3794,3795..3817,3818..3836,
                     3837..3859,3860..3878,3879..3901,3902..3920,3921..3943,
                     3944..3962,3963..3985,3986..4004)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        4019..4057
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             4181..4459
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     CDS             4193..4465
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQL"
     CDS             4472..4495
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    4823..4870
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(5957..5974)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5957..5973)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5966..5988)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6182..6201)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6301..6323
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6362..6380)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6448..6552
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6553..6621,6622..7413)
                     /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     complement(6771..6790)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7584..8172
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8073..8092
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8326..8343
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8460..8481
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8496..8501,8502..8519,8520..8526)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8534..8550
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     8539..8561
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8558..8574
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8558..8574
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 caagctagct ttgctcttag gagtttccta atacatccca aactcaaata tataaagcat
       61 ttgacttgtt ctatgcccta gggggcgggg ggaagctaag ccagcttttt ttaacattta
      121 aaatgttaat tccattttaa atgcacagat gtttttattt cataagggtt tcaatgtgca
      181 tgaatgctgc aatattcctg ttaccaaagc tagtataaat aaaaatagat aaacgtggaa
      241 attacttaga gtttctgtca ttaacgtttc cttcctcagt tgacaacata aatgcgctgc
      301 tgagcaagcc agtttgcatc tgtcaggatc aatttcccat tatgccagtc atattaatta
      361 ctagtcaatt agttgatttt tatttttgac atatacatgt gaatgaaaga ccccacctgt
      421 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg
      481 agaatagaaa agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac
      541 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct
      601 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      661 acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt
      721 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      781 gcttctcgct tctgttcgcg cgcttatgct ccccgagctc aataaaagag cccacaaccc
      841 ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata
      901 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
      961 agtgattgac tacccgtcag cgggggtctt tcatttgggg gctcgtccgg gatcgggaga
     1021 cccctgccca gggaccaccg acccaccacc gggaggtaag ctggccagca acttatctgt
     1081 gtctgtccga ttgtctagtg tctatgactg attttatgcg cctgcgtcgg tactagttag
     1141 ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttcgga acacccggcc
     1201 gcaaccctgg gagacgtccc agggacttcg ggggccgttt ttgtggcccg acctgagtcc
     1261 taaaatcccg atcgtttagg actctttggt gcacccccct tagaggaggg atatgtggtt
     1321 ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt
     1381 tgggaccgaa gccgcgccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt
     1441 ctgactgtgt ttctgtattt gtctgaaaat atgggcccgg gctagcctgt taccactccc
     1501 ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta
     1561 gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc
     1621 ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc
     1681 ttttcacctg gcccgcatgg acacccagac caggtggggt acatcgtgac ctgggaagcc
     1741 ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct
     1801 cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc
     1861 tccctttatc cagccctcac tccttctcta ggcgccccca tatggccata tgagatccga
     1921 agttcctatt ccgaagttcc tattcttcaa atagtatagg aacttcagat cataatcagc
     1981 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2041 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2101 tacaaataag gcaatagcat cacaaatttc acaaataagg catttttttc actgcattct
     2161 agttttggtt tgtccaaact catcaatgta tcttatcatg tctggatctc aaatccctcg
     2221 gaagctgcgc ctgtcttagg ttggagtgat acatttttat cacttttacc cgtctttgga
     2281 ttaggcagta gctctgacgg ccctcctgtc ttaggttagt gaaaaatgtc actctcttac
     2341 ccgtcattgg ctgtccagct tagctcgcag gggaggtggt ctggatccat gtttctcctg
     2401 ctcgaacagc tccagagcgg ctctcagctc tctgatcagc atggcggcca gggtgttccg
     2461 gtccaggttg atgccagcct cctgcagggt gatccagccc tggttcacca cgctctcctc
     2521 cactcttctc atggccatgt tgattccggc tccgatcacg atctgagcgg catcgcctgt
     2581 cttgcctgtc agctccacca ggatgccagc cagcttgcgg tcctggaggt acaggtcatt
     2641 aggccacttc actctcacct tgtcggctcc cagctttctc agcacctcgg ccatcacgat
     2701 tccgatcacc agactcagtc cgatggcagc agcaggtccc tgctccagtc tccagaacat
     2761 ggacaggtac aggttggctc cgaatgggct gaaccacttc cgtcctctgc ctcctctgcc
     2821 agcctgctgg tactcggcga tgcaggcgtc gccggacttc agctctccga ttctgtccag
     2881 caggtactga ttggtggagt cgatcacagg cagcacggcc acgcttccgc catccagctg
     2941 tcccaggatc tgcttggcgt tcagcagctg gataggctca ggcaggctgt agcccttgcc
     3001 aggcacggtg aacacgtcca ctccccagtc tctcagtgtc tggatgtgct tgttgatggc
     3061 ggctctgctc attcccaggg tctctcccag ctgctcgcca gagtggaact cgccgttggc
     3121 cagcagggcg atcagcttca ggggcacggt gttgtccttg tcgagatcct cttctgagat
     3181 gagtttttgt tcatgtttgc cccgggaggc gtaggagatc tggccgccgc cggaggagcc
     3241 gccgccacgc gtttccagtt ttagaagctc cacatcgaag acgagagtgg catgtggtgg
     3301 gatgatgcct gggtgcccag tggcaccata ggcataatct ggagatatag tcagtttggc
     3361 tctctgaccc acactcatct gggcaacccc ttcttcccag cctcggatca cctcctgctt
     3421 gcctagcata aacttaaagg gcttgtttct gtcccgggag gaatcaaatt tctttccatc
     3481 ttcaagcatc ccggtgtagt gcaccacgca ggtctggccg cgcttgggga aggtgcgccc
     3541 gtctcctggg gagatggttt ccacctgcac tcccatatcg attagctcga ccgcggaggc
     3601 tggatcggtc ccggtgtctt ctatggaggt caaaacagcg tggatggcgt ctccaggcga
     3661 tctgacggtt cactaaacga gctctgctta tataggcctc ccaccgtaca cgcctacaag
     3721 cttctttcac ttttctctat cactgatagg gagtggtaaa ctcgactttc acttttctct
     3781 atcactgata gggagtggta aactcgactt tcacttttct ctatcactga tagggagtgg
     3841 taaactcgac tttcactttt ctctatcact gatagggagt ggtaaactcg actttcactt
     3901 ttctctatca ctgataggga gtggtaaact cgactttcac ttttctctat cactgatagg
     3961 gagtggtaaa ctcgactttc acttttctct atcactgata gggagtggta aactcgaggg
     4021 taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc gtcagatcgc
     4081 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
     4141 ccgcggaatt cggccggcct caattgtttg tttaaacatg atcctctggc atgagatgtg
     4201 gcatgaaggc ctggaagagg catctcgttt gtactttggg gaaaggaacg tgaaaggcat
     4261 gtttgaggtg ctggagccct tgcatgctat gatggaacgg ggcccccaga ctctgaagga
     4321 aacatccttt aatcaggcct atggtcgaga tttaatggag gcccaagagt ggtgcaggaa
     4381 gtacatgaaa tcagggaatg tcaaggacct cacccaagcc tgggacctct attatcatgt
     4441 gttccgacga atctcaaagc agttaattaa cgactacaag gacgacgatg acaagcagat
     4501 ctcctacgcc tcccggggcg gcggctcctc cggcggcggc attaacggac tggctcctta
     4561 cctgagcaga tgggagaagc tggacaactt catcaacaga cctgtgaagc tgatcatcgg
     4621 cgacaaggaa atcttcggca tctccagagg aatcgacaag cagggagctc tgctgctgga
     4681 gcaggacgga atcatcaagc cctggatggg cggagaaatc tccctgagaa gcgcagagaa
     4741 gctcgagcgg ccgccactgt gctgtcgcga acgccgcgcg atggaacgcg cgaccctgcc
     4801 gtaataacga ggatctggcc gcgaagttcc tattccgaag ttcctattct ctagaaagta
     4861 taggaacttc agatccggat tagtccaatt tgttaaagac aggatatcag tggtccaggc
     4921 tctagttttg actcaacaat atcaccagct gaagcctata gagtacgagc catagataaa
     4981 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     5041 tggcaagcta gctcgacttc tgttcgcgcg cttctgctcc ccgagctcga gcccacaacc
     5101 cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat
     5161 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     5221 gagtgattga ctacccgtca gcgggggtct ttcacacatg cagcatgtat caaaattaat
     5281 ttggtttttt ttcttaagta tttacattaa atggccatag tacttaaagt tacattggct
     5341 tccttgaaat aaacatggag tattcagaat gtgtcataaa tatttctaat tttaagatag
     5401 tatctccatt ggctttctac tttttctttt attttttttt gtcctctgtc ttccatttgt
     5461 tgttgttgtt gtttgtttgt ttgtttgttg gttggttggt taattttttt ttaaagatcc
     5521 tacactatag ttcaagctag actattagct actctgtaac ccagggtgac cttgaagtca
     5581 tgggtagcct gctgttttag ccttcccaca tctaagatta caggtatgag ctatcatttt
     5641 tggtatattg attgattgat tgattgatgt gtgtgtgtgt gattgtgttt gtgtgtgtga
     5701 ctgtgaaaat gtgtgtatgg gtgtgtgtga atgtgtgtat gtatgtgtgt gtgtgagtgt
     5761 gtgtgtgtgt gtgtgcatgt gtgtgtgtgt gactgtgtct atgtgtatga ctgtgtgtgt
     5821 gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtgttg tgaaaaaata ttctatggta
     5881 gtgagagcca acgctccggc tcaggtgtca ggttggtttt tgagacagag tctttcactt
     5941 agcttggaat taattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     6001 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     6061 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga
     6121 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     6181 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     6241 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     6301 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg atgacgaaag
     6361 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     6421 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6481 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6541 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6601 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6661 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6721 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6781 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6841 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6901 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6961 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     7021 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     7081 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     7141 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     7201 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     7261 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     7321 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     7381 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     7441 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     7501 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     7561 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     7621 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7681 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     7741 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7801 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7861 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7921 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga
     7981 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     8041 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     8101 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     8161 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     8221 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     8281 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     8341 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     8401 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     8461 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     8521 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     8581 tacgc
//
