LOCUS       sequence-182212-        8387 bp    DNA     linear   UNA 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      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8387)
  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 8387)
  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..8387
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    complement(828..875)
                     /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)."
     CDS             complement(1203..1226)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(1233..1505)
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQL"
     CDS             complement(1239..1517)
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     promoter        complement(1641..1679)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    join(1694..1712,1713..1735,1736..1754,1755..1777,1778..1796,
                     1797..1819,1820..1838,1839..1861,1862..1880,1881..1903,
                     1904..1922,1923..1945,1946..1964)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        1980..2017
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             join(2121..2123,2124..2444)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             2505..2534
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    3489..3623
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3567..3586
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(3731..3764)
                     /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)"
     primer_bind     complement(3817..3833)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3849..3871)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    4282..4639
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             4704..5173
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5703..5719)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5703..5719)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5716..5738)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5727..5743
                     /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)."
     promoter        complement(join(5751..5757,5758..5775,5776..5781))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5796..5817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5934..5951)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6105..6693)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6185..6204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6864..7655,7656..7724))
                     /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     7487..7506
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7725..7829)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7897..7915
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7953..7975)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8075..8094
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8288..8310
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8302..8319
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8303..8319
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cacacacaca aacacaatca cacacacaca catcaatcaa tcaatcaatc aatataccaa
       61 aaatgatagc tcatacctgt aatcttagat gtgggaaggc taaaacagca ggctacccat
      121 gacttcaagg tcaccctggg ttacagagta gctaatagtc tagcttgaac tatagtgtag
      181 gatctttaaa aaaaaattaa ccaaccaacc aacaaacaaa caaacaaaca acaacaacaa
      241 caaatggaag acagaggaca aaaaaaaata aaagaaaaag tagaaagcca atggagatac
      301 tatcttaaaa ttagaaatat ttatgacaca ttctgaatac tccatgttta tttcaaggaa
      361 gccaatgtaa ctttaagtac tatggccatt taatgtaaat acttaagaaa aaaaaccaaa
      421 ttaattttga tacatgctgc atgtgaaaga cccccgctga cgggtagtca atcactcaga
      481 ggagaccctc ccaaggaaca gcgagaccac aagtcggatg caactgcaag agggtttatt
      541 ggatacacgg gtacccgggc gactcagtca atcggaggac tggcgccccg agtgaggggt
      601 tgtgggctcg agctcgggga gcagaagcgc gcgaacagaa gtcgagctag cttgccaaac
      661 ctacaggtgg ggtctttcat tccccccttt ttctggagac taaataaaat cttttatttt
      721 atctatggct cgtactctat aggcttcagc tggtgatatt gttgagtcaa aactagagcc
      781 tggaccactg atatcctgtc tttaacaaat tggactaatc cggatctgaa gttcctatac
      841 tttctagaga ataggaactt cggaatagga acttcgcggc cagatcctcg ttattacggc
      901 agggtcgcgc gttccatcgc gcggcgttcg cgacagcaca gtggcggccg ctcgagcttc
      961 tctgcgcttc tcagggagat ttctccgccc atccagggct tgatgattcc gtcctgctcc
     1021 agcagcagag ctccctgctt gtcgattcct ctggagatgc cgaagatttc cttgtcgccg
     1081 atgatcagct tcacaggtct gttgatgaag ttgtccagct tctcccatct gctcaggtaa
     1141 ggagccagtc cgttaatgcc gccgccggag gagccgccgc cccgggaggc gtaggagatc
     1201 tgcttgtcat cgtcgtcctt gtagtcgtta attaactgct ttgagattcg tcggaacaca
     1261 tgataataga ggtcccaggc ttgggtgagg tccttgacat tccctgattt catgtacttc
     1321 ctgcaccact cttgggcctc cattaaatct cgaccatagg cctgattaaa ggatgtttcc
     1381 ttcagagtct gggggccccg ttccatcata gcatgcaagg gctccagcac ctcaaacatg
     1441 cctttcacgt tcctttcccc aaagtacaaa cgagatgcct cttccaggcc ttcatgccac
     1501 atctcatgcc agaggatcat gtttaaacaa acaattgagg ccggccgaat tccgcggagg
     1561 ctggatcggt cccggtgtct tctatggagg tcaaaacagc gtggatggcg tctccaggcg
     1621 atctgacggt tcactaaacg agctctgctt atataggcct cccaccgtac acgcctaccc
     1681 tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct
     1741 atcagtgata gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag
     1801 tgaaagtcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca
     1861 ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga
     1921 gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg aaaaagcttg
     1981 taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc gtcagatcgc
     2041 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
     2101 ccgcggtcga gctaatcgat atgggagtgc aggtggaaac catctcccca ggagacgggc
     2161 gcaccttccc caagcgcggc cagacctgcg tggtgcacta caccgggatg cttgaagatg
     2221 gaaagaaatt tgattcctcc cgggacagaa acaagccctt taagtttatg ctaggcaagc
     2281 aggaggtgat ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt cagagagcca
     2341 aactgactat atctccagat tatgcctatg gtgccactgg gcacccaggc atcatcccac
     2401 cacatgccac tctcgtcttc gatgtggagc ttctaaaact ggaaacgcgt ggcggcggct
     2461 cctccggcgg cggccagatc tcctacgcct cccggggcaa acatgaacaa aaactcatct
     2521 cagaagagga tctcgacaag gacaacaccg tgcccctgaa gctgatcgcc ctgctggcca
     2581 acggcgagtt ccactctggc gagcagctgg gagagaccct gggaatgagc agagccgcca
     2641 tcaacaagca catccagaca ctgagagact ggggagtgga cgtgttcacc gtgcctggca
     2701 agggctacag cctgcctgag cctatccagc tgctgaacgc caagcagatc ctgggacagc
     2761 tggatggcgg aagcgtggcc gtgctgcctg tgatcgactc caccaatcag tacctgctgg
     2821 acagaatcgg agagctgaag tccggcgacg cctgcatcgc cgagtaccag caggctggca
     2881 gaggaggcag aggacggaag tggttcagcc cattcggagc caacctgtac ctgtccatgt
     2941 tctggagact ggagcaggga cctgctgctg ccatcggact gagtctggtg atcggaatcg
     3001 tgatggccga ggtgctgaga aagctgggag ccgacaaggt gagagtgaag tggcctaatg
     3061 acctgtacct ccaggaccgc aagctggctg gcatcctggt ggagctgaca ggcaagacag
     3121 gcgatgccgc tcagatcgtg atcggagccg gaatcaacat ggccatgaga agagtggagg
     3181 agagcgtggt gaaccagggc tggatcaccc tgcaggaggc tggcatcaac ctggaccgga
     3241 acaccctggc cgccatgctg atcagagagc tgagagccgc tctggagctg ttcgagcagg
     3301 agaaacatgg atccagacca cctcccctgc gagctaagct ggacagccaa tgacgggtaa
     3361 gagagtgaca tttttcacta acctaagaca ggagggccgt cagagctact gcctaatcca
     3421 aagacgggta aaagtgataa aaatgtatca ctccaaccta agacaggcgc agcttccgag
     3481 ggatttgaga tccagacatg ataagataca ttgatgagtt tggacaaacc aaaactagaa
     3541 tgcagtgaaa aaaatgcctt atttgtgaaa tttgtgatgc tattgcctta tttgtaacca
     3601 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     3661 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
     3721 attatgatct gaagttccta tactatttga agaataggaa cttcggaata ggaacttcgg
     3781 atctcatatg gccatatggg ggcgcctaga gaaggagtga gggctggata aagggaggat
     3841 cgaggcgggg tcgaacgagg aggttcaagg gggagagacg gggcggatgg aggaagagga
     3901 ggcggaggct tagggtgtac aaagggcttg acccagggag gggggtcaaa agccaaggct
     3961 tcccaggtca cgatgtaggg gacctggtct gggtgtccat gcgggccagg tgaaaagacc
     4021 ttgatcttaa cctgggtgat gaggtctcgg ttaaaggtgc cgtctcgcgg ccatccgacg
     4081 ttaaaggttg gccattctgc agagcagaag gtaacccaac gtctcttctt gacatctacc
     4141 gactggttgt gagcgatccg ctcgacatct ttccagtgac ctaaggtcaa acttaaggga
     4201 gtggtaacag gctagcccgg gcccatattt tcagacaaat acagaaacac agtcagacag
     4261 agacaacaca gaacgatgct gcagcagaca agacgcgcgg cgcggcttcg gtcccaaacc
     4321 gaaagcaaaa attcagacgg aggcgggaac tgttttaggt tctcgtctcc taccagaacc
     4381 acatatccct cctctaaggg gggtgcacca aagagtccta aacgatcggg attttaggac
     4441 tcaggtcggg ccacaaaaac ggcccccgaa gtccctggga cgtctcccag ggttgcggcc
     4501 gggtgttccg aactcgtcag ttccaccacg ggtccgccag atacagagct agttagctaa
     4561 ctagtaccga cgcaggcgca taaaatcagt catagacact agacaatcgg acagacacag
     4621 ataagttgct ggccagctta cctcccggtg gtgggtcggt ggtccctggg caggggtctc
     4681 ccgatcccgg acgagccccc aaatgaaaga cccccgctga cgggtagtca atcactcaga
     4741 ggagaccctc ccaaggaaca gcgagaccac aagtcggatg caactgcaag agggtttatt
     4801 ggatacacgg gtacccgggc gactcagtca atcggaggac tggcgcgccg agtgaggggt
     4861 tgtgggctct tttattgagc tcggggagca gaagcgcgcg aacagaagcg agaagcgaac
     4921 tgattggtta gttcaaataa ggcacagggt catttcaggt ccttggggca ccctggaaac
     4981 atctgatggt tctctagaaa ctgctgaggg ctggaccgca tctggggacc atctgttctt
     5041 ggccctgagc cggggcagga actgcttacc acagatatcc tgtttggccc atattcagct
     5101 gttccatctg ttcttggccc tgagccgggg caggaactgc ttaccacaga tatcctgttt
     5161 ggcccatatt cagctgttcc atctgttcct gaccttgatc tgaacttttc tattctcagt
     5221 tatgtatttt tccatgcctt gcaaaatggc gttacttaag ctagcttgcc aaacctacag
     5281 gtggggtctt tcacatgtat atgtcaaaaa taaaaatcaa ctaattgact agtaattaat
     5341 atgactggca taatgggaaa ttgatcctga cagatgcaaa ctggcttctc agcagcgcat
     5401 ttatgttgtc aactgaggaa ggaaacgtta atgacagaaa ctctaagtaa tttccacgtt
     5461 tatctatttt tatttatact agctttggta acaggaatat tgcagcattc atgcacattg
     5521 aaacccttat gaaataaaaa catctgtgca tttaaaatgg aattaacatt ttaaatgtta
     5581 aaaaaagctg gcttagcttc cccccgcccc ctagggcata gaacaagtca aatgctttat
     5641 atatttgagt ttgggatgta ttaggaaact cctaagagca aagctagctt ggcgtaatca
     5701 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5761 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5821 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5881 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5941 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6001 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6061 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6121 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6181 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6241 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6301 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6361 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6421 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6481 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6541 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6601 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6661 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6721 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6781 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6841 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6901 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6961 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7021 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7081 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7141 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7201 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7261 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7321 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7381 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7441 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7501 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7561 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7621 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7681 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7741 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7801 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7861 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     7921 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     7981 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     8041 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     8101 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     8161 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     8221 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     8281 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattaattcc aagctaagtg
     8341 aaagactctg tctcaaaaac caacctgaca cctgagccgg agcgttg
//
