LOCUS       sequence-177762-        8582 bp    DNA     circular SYN 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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8582)
  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 8582)
  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..8582
                     /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(join(2398..2718,2719..2721))
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             complement(3550..3579)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(3677..3714)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    join(3731..3749,3750..3772,3773..3791,3792..3814,3815..3833,
                     3834..3856,3857..3875,3876..3898,3899..3917,3918..3940,
                     3941..3959,3960..3982,3983..4001)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        4016..4054
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             4178..4456
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     CDS             4190..4462
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQL"
     CDS             4469..4492
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    4820..4867
                     /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(5954..5971)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5954..5970)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5963..5985)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6179..6198)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6298..6320
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6359..6377)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6445..6549
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6550..6618,6619..7410)
                     /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(6768..6787)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7581..8169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8070..8089
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8323..8340
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8457..8478
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8493..8498,8499..8516,8517..8523)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8531..8547
                     /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     8536..8558
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8555..8571
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8555..8571
                     /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 ctggatccct aacgcgtttc
     2401 cagttttaga agctccacat cgaagacgag agtggcatgt ggtgggatga tgcctgggtg
     2461 cccagtggca ccataggcat aatctggaga tatagtcagt ttggctctct gacccacact
     2521 catctgggca accccttctt cccagcctcg gatcacctcc tgcttgccta gcataaactt
     2581 aaagggcttg tttctgtccc gggaggaatc aaatttcttt ccatcttcaa gcatcccggt
     2641 gtagtgcacc acgcaggtct ggccgcgctt ggggaaggtg cgcccgtctc ctggggagat
     2701 ggtttccacc tgcactccca tatcgatgcc ccgggaggcg taggagatct ggccgccgcc
     2761 ggaggagccg ccgccatgtt tctcctgctc gaacagctcc agagcggctc tcagctctct
     2821 gatcagcatg gcggccaggg tgttccggtc caggttgatg ccagcctcct gcagggtgat
     2881 ccagccctgg ttcaccacgc tctcctccac tcttctcatg gccatgttga ttccggctcc
     2941 gatcacgatc tgagcggcat cgcctgtctt gcctgtcagc tccaccagga tgccagccag
     3001 cttgcggtcc tggaggtaca ggtcattagg ccacttcact ctcaccttgt cggctcccag
     3061 ctttctcagc acctcggcca tcacgattcc gatcaccaga ctcagtccga tggcagcagc
     3121 aggtccctgc tccagtctcc agaacatgga caggtacagg ttggctccga atgggctgaa
     3181 ccacttccgt cctctgcctc ctctgccagc ctgctggtac tcggcgatgc aggcgtcgcc
     3241 ggacttcagc tctccgattc tgtccagcag gtactgattg gtggagtcga tcacaggcag
     3301 cacggccacg cttccgccat ccagctgtcc caggatctgc ttggcgttca gcagctggat
     3361 aggctcaggc aggctgtagc ccttgccagg cacggtgaac acgtccactc cccagtctct
     3421 cagtgtctgg atgtgcttgt tgatggcggc tctgctcatt cccagggtct ctcccagctg
     3481 ctcgccagag tggaactcgc cgttggccag cagggcgatc agcttcaggg gcacggtgtt
     3541 gtccttgtcg agatcctctt ctgagatgag tttttgttcc atctcgaccg cggaggctgg
     3601 atcggtcccg gtgtcttcta tggaggtcaa aacagcgtgg atggcgtctc caggcgatct
     3661 gacggttcac taaacgagct ctgcttatat aggcctccca ccgtacacgc ctacaagctt
     3721 ctttcacttt tctctatcac tgatagggag tggtaaactc gactttcact tttctctatc
     3781 actgataggg agtggtaaac tcgactttca cttttctcta tcactgatag ggagtggtaa
     3841 actcgacttt cacttttctc tatcactgat agggagtggt aaactcgact ttcacttttc
     3901 tctatcactg atagggagtg gtaaactcga ctttcacttt tctctatcac tgatagggag
     3961 tggtaaactc gactttcact tttctctatc actgataggg agtggtaaac tcgagggtag
     4021 gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     4081 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg
     4141 cggaattcgg ccggcctcaa ttgtttgttt aaacatgatc ctctggcatg agatgtggca
     4201 tgaaggcctg gaagaggcat ctcgtttgta ctttggggaa aggaacgtga aaggcatgtt
     4261 tgaggtgctg gagcccttgc atgctatgat ggaacggggc ccccagactc tgaaggaaac
     4321 atcctttaat caggcctatg gtcgagattt aatggaggcc caagagtggt gcaggaagta
     4381 catgaaatca gggaatgtca aggacctcac ccaagcctgg gacctctatt atcatgtgtt
     4441 ccgacgaatc tcaaagcagt taattaacga ctacaaggac gacgatgaca agcagatctc
     4501 ctacgcctcc cggggcggcg gctcctccgg cggcggcatt aacggactgg ctccttacct
     4561 gagcagatgg gagaagctgg acaacttcat caacagacct gtgaagctga tcatcggcga
     4621 caaggaaatc ttcggcatct ccagaggaat cgacaagcag ggagctctgc tgctggagca
     4681 ggacggaatc atcaagccct ggatgggcgg agaaatctcc ctgagaagcg cagagaagct
     4741 cgagcggccg ccactgtgct gtcgcgaacg ccgcgcgatg gaacgcgcga ccctgccgta
     4801 ataacgagga tctggccgcg aagttcctat tccgaagttc ctattctcta gaaagtatag
     4861 gaacttcaga tccggattag tccaatttgt taaagacagg atatcagtgg tccaggctct
     4921 agttttgact caacaatatc accagctgaa gcctatagag tacgagccat agataaaata
     4981 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     5041 caagctagct cgacttctgt tcgcgcgctt ctgctccccg agctcgagcc cacaacccct
     5101 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     5161 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     5221 tgattgacta cccgtcagcg ggggtctttc acacatgcag catgtatcaa aattaatttg
     5281 gttttttttc ttaagtattt acattaaatg gccatagtac ttaaagttac attggcttcc
     5341 ttgaaataaa catggagtat tcagaatgtg tcataaatat ttctaatttt aagatagtat
     5401 ctccattggc tttctacttt ttcttttatt tttttttgtc ctctgtcttc catttgttgt
     5461 tgttgttgtt tgtttgtttg tttgttggtt ggttggttaa ttttttttta aagatcctac
     5521 actatagttc aagctagact attagctact ctgtaaccca gggtgacctt gaagtcatgg
     5581 gtagcctgct gttttagcct tcccacatct aagattacag gtatgagcta tcatttttgg
     5641 tatattgatt gattgattga ttgatgtgtg tgtgtgtgat tgtgtttgtg tgtgtgactg
     5701 tgaaaatgtg tgtatgggtg tgtgtgaatg tgtgtatgta tgtgtgtgtg tgagtgtgtg
     5761 tgtgtgtgtg tgcatgtgtg tgtgtgtgac tgtgtctatg tgtatgactg tgtgtgtgtg
     5821 tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgttgtga aaaaatattc tatggtagtg
     5881 agagccaacg ctccggctca ggtgtcaggt tggtttttga gacagagtct ttcacttagc
     5941 ttggaattaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     6001 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     6061 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     6121 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
     6181 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     6241 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     6301 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgatg acgaaagggc
     6361 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     6421 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6481 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6541 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6601 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6661 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6721 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6781 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6841 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6901 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6961 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     7021 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     7081 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     7141 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     7201 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7261 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7321 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7381 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7441 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7501 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7561 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7621 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7681 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     7741 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7801 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7861 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7921 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa
     7981 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     8041 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     8101 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     8161 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     8221 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     8281 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     8341 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     8401 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     8461 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     8521 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     8581 gc
//
