LOCUS       sequence-182190-        8573 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Split-BioID plasmid: Expresses C-terminally tagged FRB-CBir[257-321]
            and N-terminally tagged NBir[2-256]-FKBP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8573)
  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 8573)
  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_Myc-NBir-FKBP
FEATURES             Location/Qualifiers
     source          1..8573
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    complement(1016..1063)
                     /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(1391..1414)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(1421..1693)
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQL"
     CDS             complement(1427..1705)
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     promoter        complement(1829..1867)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    join(1882..1900,1901..1923,1924..1942,1943..1965,1966..1984,
                     1985..2007,2008..2026,2027..2049,2050..2068,2069..2091,
                     2092..2110,2111..2133,2134..2152)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2169..2206
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             2304..2333
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(3162..3164,3165..3485)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     polyA_signal    3675..3809
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3753..3772
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(3917..3950)
                     /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(4003..4019)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(4035..4057)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    4468..4825
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             4890..5359
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5889..5905)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5889..5905)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5902..5924)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5913..5929
                     /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(5937..5943,5944..5961,5962..5967))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5982..6003
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6120..6137)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6291..6879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6371..6390)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7050..7841,7842..7910))
                     /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     7673..7692
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7911..8015)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8083..8101
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8139..8161)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8261..8280
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8474..8496
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8488..8505
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8489..8505
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cctctcacta ccataaatat atatacacaa cacacacaca cacacacaca cacacacaca
       61 cacacacaca cacacaatca tacacataaa cacaatcaca cacacacaca tgcacaatca
      121 cacacacaca atcacacaca aacatacata cacacaatca cacacaacca tacacacata
      181 tacacaatca cacacacaaa cacaatcaca cacacacaca tcaatcaatc aatcaatcaa
      241 tataccaaaa atgatagctc atacctgtaa tcttagatgt gggaaggcta aaacagcagg
      301 ctacccatga cttcaaggtc accctgggtt acagagtagc taatagtcta gcttgaacta
      361 tagtgtagga tctttaaaaa aaaattaacc aaccaaccaa caaacaaaca aacaaacaac
      421 aacaacaaca aatggaagac agaggacaaa aaaaaataaa agaaaaagta gaaagccaat
      481 ggagatacta tcttaaaatt agaaatattt atgacacatt ctgaatactc catgtttatt
      541 tcaaggaagc caatgtaact ttaagtacta tggccattta atgtaaatac ttaagaaaaa
      601 aaaccaaatt aattttgata catgctgcat gtgaaagacc cccgctgacg ggtagtcaat
      661 cactcagagg agaccctccc aaggaacagc gagaccacaa gtcggatgca actgcaagag
      721 ggtttattgg atacacgggt acccgggcga ctcagtcaat cggaggactg gcgccccgag
      781 tgaggggttg tgggctcgag ctcggggagc agaagcgcgc gaacagaagt cgagctagct
      841 tgccaaacct acaggtgggg tctttcattc cccccttttt ctggagacta aataaaatct
      901 tttattttat ctatggctcg tactctatag gcttcagctg gtgatattgt tgagtcaaaa
      961 ctagagcctg gaccactgat atcctgtctt taacaaattg gactaatccg gatctgaagt
     1021 tcctatactt tctagagaat aggaacttcg gaataggaac ttcgcggcca gatcctcgtt
     1081 attacggcag ggtcgcgcgt tccatcgcgc ggcgttcgcg acagcacagt ggcggccgct
     1141 cgagcttctc tgcgcttctc agggagattt ctccgcccat ccagggcttg atgattccgt
     1201 cctgctccag cagcagagct ccctgcttgt cgattcctct ggagatgccg aagatttcct
     1261 tgtcgccgat gatcagcttc acaggtctgt tgatgaagtt gtccagcttc tcccatctgc
     1321 tcaggtaagg agccagtccg ttaatgccgc cgccggagga gccgccgccc cgggaggcgt
     1381 aggagatctg cttgtcatcg tcgtccttgt agtcgttaat taactgcttt gagattcgtc
     1441 ggaacacatg ataatagagg tcccaggctt gggtgaggtc cttgacattc cctgatttca
     1501 tgtacttcct gcaccactct tgggcctcca ttaaatctcg accataggcc tgattaaagg
     1561 atgtttcctt cagagtctgg gggccccgtt ccatcatagc atgcaagggc tccagcacct
     1621 caaacatgcc tttcacgttc ctttccccaa agtacaaacg agatgcctct tccaggcctt
     1681 catgccacat ctcatgccag aggatcatgt ttaaacaaac aattgaggcc ggccgaattc
     1741 cgcggaggct ggatcggtcc cggtgtcttc tatggaggtc aaaacagcgt ggatggcgtc
     1801 tccaggcgat ctgacggttc actaaacgag ctctgcttat ataggcctcc caccgtacac
     1861 gcctaccctc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
     1921 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
     1981 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
     2041 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
     2101 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
     2161 agaagcttgt aggcgtgtac ggtgggaggc ctatataagc agagctcgtt tagtgaaccg
     2221 tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg
     2281 atccagcctc cgcggtcgag atggaacaaa aactcatctc agaagaggat ctcgacaagg
     2341 acaacaccgt gcccctgaag ctgatcgccc tgctggccaa cggcgagttc cactctggcg
     2401 agcagctggg agagaccctg ggaatgagca gagccgccat caacaagcac atccagacac
     2461 tgagagactg gggagtggac gtgttcaccg tgcctggcaa gggctacagc ctgcctgagc
     2521 ctatccagct gctgaacgcc aagcagatcc tgggacagct ggatggcgga agcgtggccg
     2581 tgctgcctgt gatcgactcc accaatcagt acctgctgga cagaatcgga gagctgaagt
     2641 ccggcgacgc ctgcatcgcc gagtaccagc aggctggcag aggaggcaga ggacggaagt
     2701 ggttcagccc attcggagcc aacctgtacc tgtccatgtt ctggagactg gagcagggac
     2761 ctgctgctgc catcggactg agtctggtga tcggaatcgt gatggccgag gtgctgagaa
     2821 agctgggagc cgacaaggtg agagtgaagt ggcctaatga cctgtacctc caggaccgca
     2881 agctggctgg catcctggtg gagctgacag gcaagacagg cgatgccgct cagatcgtga
     2941 tcggagccgg aatcaacatg gccatgagaa gagtggagga gagcgtggtg aaccagggct
     3001 ggatcaccct gcaggaggct ggcatcaacc tggaccggaa caccctggcc gccatgctga
     3061 tcagagagct gagagccgct ctggagctgt tcgagcagga gaaacatggc ggcggctcct
     3121 ccggcggcgg ccagatctcc tacgcctccc ggggcatcga tatgggagtg caggtggaaa
     3181 ccatctcccc aggagacggg cgcaccttcc ccaagcgcgg ccagacctgc gtggtgcact
     3241 acaccgggat gcttgaagat ggaaagaaat ttgattcctc ccgggacaga aacaagccct
     3301 ttaagtttat gctaggcaag caggaggtga tccgaggctg ggaagaaggg gttgcccaga
     3361 tgagtgtggg tcagagagcc aaactgacta tatctccaga ttatgcctat ggtgccactg
     3421 ggcacccagg catcatccca ccacatgcca ctctcgtctt cgatgtggag cttctaaaac
     3481 tggaaacgcg ttagggatcc agaccacctc ccctgcgagc taagctggac agccaatgac
     3541 gggtaagaga gtgacatttt tcactaacct aagacaggag ggccgtcaga gctactgcct
     3601 aatccaaaga cgggtaaaag tgataaaaat gtatcactcc aacctaagac aggcgcagct
     3661 tccgagggat ttgagatcca gacatgataa gatacattga tgagtttgga caaaccaaaa
     3721 ctagaatgca gtgaaaaaaa tgccttattt gtgaaatttg tgatgctatt gccttatttg
     3781 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3841 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     3901 tggctgatta tgatctgaag ttcctatact atttgaagaa taggaacttc ggaataggaa
     3961 cttcggatct catatggcca tatgggggcg cctagagaag gagtgagggc tggataaagg
     4021 gaggatcgag gcggggtcga acgaggaggt tcaaggggga gagacggggc ggatggagga
     4081 agaggaggcg gaggcttagg gtgtacaaag ggcttgaccc agggaggggg gtcaaaagcc
     4141 aaggcttccc aggtcacgat gtaggggacc tggtctgggt gtccatgcgg gccaggtgaa
     4201 aagaccttga tcttaacctg ggtgatgagg tctcggttaa aggtgccgtc tcgcggccat
     4261 ccgacgttaa aggttggcca ttctgcagag cagaaggtaa cccaacgtct cttcttgaca
     4321 tctaccgact ggttgtgagc gatccgctcg acatctttcc agtgacctaa ggtcaaactt
     4381 aagggagtgg taacaggcta gcccgggccc atattttcag acaaatacag aaacacagtc
     4441 agacagagac aacacagaac gatgctgcag cagacaagac gcgcggcgcg gcttcggtcc
     4501 caaaccgaaa gcaaaaattc agacggaggc gggaactgtt ttaggttctc gtctcctacc
     4561 agaaccacat atccctcctc taaggggggt gcaccaaaga gtcctaaacg atcgggattt
     4621 taggactcag gtcgggccac aaaaacggcc cccgaagtcc ctgggacgtc tcccagggtt
     4681 gcggccgggt gttccgaact cgtcagttcc accacgggtc cgccagatac agagctagtt
     4741 agctaactag taccgacgca ggcgcataaa atcagtcata gacactagac aatcggacag
     4801 acacagataa gttgctggcc agcttacctc ccggtggtgg gtcggtggtc cctgggcagg
     4861 ggtctcccga tcccggacga gcccccaaat gaaagacccc cgctgacggg tagtcaatca
     4921 ctcagaggag accctcccaa ggaacagcga gaccacaagt cggatgcaac tgcaagaggg
     4981 tttattggat acacgggtac ccgggcgact cagtcaatcg gaggactggc gcgccgagtg
     5041 aggggttgtg ggctctttta ttgagctcgg ggagcagaag cgcgcgaaca gaagcgagaa
     5101 gcgaactgat tggttagttc aaataaggca cagggtcatt tcaggtcctt ggggcaccct
     5161 ggaaacatct gatggttctc tagaaactgc tgagggctgg accgcatctg gggaccatct
     5221 gttcttggcc ctgagccggg gcaggaactg cttaccacag atatcctgtt tggcccatat
     5281 tcagctgttc catctgttct tggccctgag ccggggcagg aactgcttac cacagatatc
     5341 ctgtttggcc catattcagc tgttccatct gttcctgacc ttgatctgaa cttttctatt
     5401 ctcagttatg tatttttcca tgccttgcaa aatggcgtta cttaagctag cttgccaaac
     5461 ctacaggtgg ggtctttcac atgtatatgt caaaaataaa aatcaactaa ttgactagta
     5521 attaatatga ctggcataat gggaaattga tcctgacaga tgcaaactgg cttctcagca
     5581 gcgcatttat gttgtcaact gaggaaggaa acgttaatga cagaaactct aagtaatttc
     5641 cacgtttatc tatttttatt tatactagct ttggtaacag gaatattgca gcattcatgc
     5701 acattgaaac ccttatgaaa taaaaacatc tgtgcattta aaatggaatt aacattttaa
     5761 atgttaaaaa aagctggctt agcttccccc cgccccctag ggcatagaac aagtcaaatg
     5821 ctttatatat ttgagtttgg gatgtattag gaaactccta agagcaaagc tagcttggcg
     5881 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5941 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6001 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6061 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6121 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6181 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6241 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6301 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6361 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6421 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6481 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6541 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6601 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6661 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6721 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6781 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6841 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6901 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6961 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7021 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7081 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7141 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7201 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7261 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7321 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7381 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7441 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7501 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7561 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7621 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7681 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7741 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7801 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7861 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7921 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7981 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8041 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     8101 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     8161 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     8221 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     8281 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     8341 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     8401 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     8461 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt aattccaagc
     8521 taagtgaaag actctgtctc aaaaaccaac ctgacacctg agccggagcg ttg
//
