LOCUS       sequence-413994-        8538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Split-BioID plasmid: Expresses N-terminally tagged CBir[257-321]-FRB
            and C-terminally tagged FKBP-NBir[2-256].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8538)
  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 8538)
  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-CBir-FRB_FKBP-Myc-Nbir
FEATURES             Location/Qualifiers
     source          1..8538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             519..988
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    1053..1410
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     primer_bind     1821..1843
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1859..1875
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    1928..1961
                     /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    2069..2203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2106..2125)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3158..3187)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(join(3248..3568,3569..3571))
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     promoter        complement(3675..3712)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    join(3729..3747,3748..3770,3771..3789,3790..3812,3813..3831,
                     3832..3854,3855..3873,3874..3896,3897..3915,3916..3938,
                     3939..3957,3958..3980,3981..3999)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        4014..4052
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             4164..4187
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4461..4739
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     CDS             4473..4745
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQL"
     protein_bind    4780..4827
                     /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(5911..5928)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5911..5927)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5920..5942)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6136..6155)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6255..6277
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6315..6333)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6401..6505
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6506..6574,6575..7366)
                     /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(6724..6743)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7537..8125
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8026..8045
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8279..8296
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8413..8434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8449..8454,8455..8472,8473..8479)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8487..8503
                     /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     8492..8514
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8511..8527
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8511..8527
                     /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 tgagaagcca gtttgcatct gtcaggatca atttcccatt atgccagtca tattaattac
      361 tagtcaatta gttgattttt atttttgaca tatacatgtg aaagacccca cctgtaggtt
      421 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat
      481 agaaaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat
      541 atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata
      601 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
      661 tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg
      721 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
      781 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
      841 tcggcgcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc
      901 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga
      961 ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg ggagacccct
     1021 gcccagggac caccgaccca ccaccgggag gtaagctggc cagcaactta tctgtgtctg
     1081 tccgattgtc tagtgtctat gactgatttt atgcgcctgc gtcggtacta gttagctaac
     1141 tagctctgta tctggcggac ccgtggtgga actgacgagt tcggaacacc cggccgcaac
     1201 cctgggagac gtcccaggga cttcgggggc cgtttttgtg gcccgacctg agtcctaaaa
     1261 tcccgatcgt ttaggactct ttggtgcacc ccccttagag gagggatatg tggttctggt
     1321 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggga
     1381 ccgaagccgc gccgcgcgtc ttgtctgctg cagcatcgtt ctgtgttgtc tctgtctgac
     1441 tgtgtttctg tatttgtctg aaaatatggg cccgggctag cctgttacca ctcccttaag
     1501 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1561 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1621 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1681 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1741 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1801 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1861 ttatccagcc ctcactcctt ctctaggcgc ccccatatgg ccatatgaga tccgaagttc
     1921 ctattccgaa gttcctattc ttcaaatagt ataggaactt cagatcataa tcagccatac
     1981 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     2041 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2101 ataaggcaat agcatcacaa atttcacaaa taaggcattt ttttcactgc attctagttt
     2161 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctcaaatc cctcggaagc
     2221 tgcgcctgtc ttaggttgga gtgatacatt tttatcactt ttacccgtct ttggattagg
     2281 cagtagctct gacggccctc ctgtcttagg ttagtgaaaa atgtcactct cttacccgtc
     2341 attggctgtc cagcttagct cgcaggggag gtggtctgga tccatgtttc tcctgctcga
     2401 acagctccag agcggctctc agctctctga tcagcatggc ggccagggtg ttccggtcca
     2461 ggttgatgcc agcctcctgc agggtgatcc agccctggtt caccacgctc tcctccactc
     2521 ttctcatggc catgttgatt ccggctccga tcacgatctg agcggcatcg cctgtcttgc
     2581 ctgtcagctc caccaggatg ccagccagct tgcggtcctg gaggtacagg tcattaggcc
     2641 acttcactct caccttgtcg gctcccagct ttctcagcac ctcggccatc acgattccga
     2701 tcaccagact cagtccgatg gcagcagcag gtccctgctc cagtctccag aacatggaca
     2761 ggtacaggtt ggctccgaat gggctgaacc acttccgtcc tctgcctcct ctgccagcct
     2821 gctggtactc ggcgatgcag gcgtcgccgg acttcagctc tccgattctg tccagcaggt
     2881 actgattggt ggagtcgatc acaggcagca cggccacgct tccgccatcc agctgtccca
     2941 ggatctgctt ggcgttcagc agctggatag gctcaggcag gctgtagccc ttgccaggca
     3001 cggtgaacac gtccactccc cagtctctca gtgtctggat gtgcttgttg atggcggctc
     3061 tgctcattcc cagggtctct cccagctgct cgccagagtg gaactcgccg ttggccagca
     3121 gggcgatcag cttcaggggc acggtgttgt ccttgtcgag atcctcttct gagatgagtt
     3181 tttgttcatg tttgccccgg gaggcgtagg agatctggcc gccgccggag gagccgccgc
     3241 cacgcgtttc cagttttaga agctccacat cgaagacgag agtggcatgt ggtgggatga
     3301 tgcctgggtg cccagtggca ccataggcat aatctggaga tatagtcagt ttggctctct
     3361 gacccacact catctgggca accccttctt cccagcctcg gatcacctcc tgcttgccta
     3421 gcataaactt aaagggcttg tttctgtccc gggaggaatc aaatttcttt ccatcttcaa
     3481 gcatcccggt gtagtgcacc acgcaggtct ggccgcgctt ggggaaggtg cgcccgtctc
     3541 ctggggagat ggtttccacc tgcactccca tatcgattag ctcgaccgcg gaggctggat
     3601 cggtcccggt gtcttctatg gaggtcaaaa cagcgtggat ggcgtctcca ggcgatctga
     3661 cggttcacta aacgagctct gcttatatag gcctcccacc gtacacgcct acaagcttct
     3721 ttcacttttc tctatcactg atagggagtg gtaaactcga ctttcacttt tctctatcac
     3781 tgatagggag tggtaaactc gactttcact tttctctatc actgataggg agtggtaaac
     3841 tcgactttca cttttctcta tcactgatag ggagtggtaa actcgacttt cacttttctc
     3901 tatcactgat agggagtggt aaactcgact ttcacttttc tctatcactg atagggagtg
     3961 gtaaactcga ctttcacttt tctctatcac tgatagggag tggtaaactc gagggtaggc
     4021 gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga
     4081 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgcg
     4141 gaattcggcc ggcctcaatt atggactaca aggacgacga tgacaagatt aacggactgg
     4201 ctccttacct gagcagatgg gagaagctgg acaacttcat caacagacct gtgaagctga
     4261 tcatcggcga caaggaaatc ttcggcatct ccagaggaat cgacaagcag ggagctctgc
     4321 tgctggagca ggacggaatc atcaagccct ggatgggcgg agaaatctcc ctgagaagcg
     4381 cagagaagct cgaggcgaac cagatctcct acgcctcccg gggcggcggc tcctccggcg
     4441 gcggcgtttg tttaaacatg atcctctggc atgagatgtg gcatgaaggc ctggaagagg
     4501 catctcgttt gtactttggg gaaaggaacg tgaaaggcat gtttgaggtg ctggagccct
     4561 tgcatgctat gatggaacgg ggcccccaga ctctgaagga aacatccttt aatcaggcct
     4621 atggtcgaga tttaatggag gcccaagagt ggtgcaggaa gtacatgaaa tcagggaatg
     4681 tcaaggacct cacccaagcc tgggacctct attatcatgt gttccgacga atctcaaagc
     4741 agttaattaa atagcgatta ataacgagga tctggccgcg aagttcctat tccgaagttc
     4801 ctattctcta gaaagtatag gaacttcaga tccggattag tccaatttgt taaagacagg
     4861 atatcagtgg tccaggctct agttttgact caacaatatc accagctgaa gcctatagag
     4921 tacgagccat agataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa
     4981 gaccccacct gtaggtttgg caagctagct cgacttctgt tcgcgcgctt ctgctccccg
     5041 agctcgagcc cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg
     5101 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
     5161 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc acatgcagca
     5221 tgtatcaaaa ttaatttggt tttttttctt aagtatttac attaaatggc catagtactt
     5281 aaagttacat tggcttcctt gaaataaaca tggagtattc agaatgtgtc ataaatattt
     5341 ctaattttaa gatagtatct ccattggctt tctacttttt cttttatttt tttttgtcct
     5401 ctgtcttcca tttgttgttg ttgttgtttg tttgtttgtt tgttggttgg ttggttaatt
     5461 tttttttaaa gatcctacac tatagttcaa gctagactat tagctactct gtaacccagg
     5521 gtgaccttga agtcatgggt agcctgctgt tttagccttc ccacatctaa gattacaggt
     5581 atgagctatc atttttggta tattgattga ttgattgatt gatgtgtgtg tgtgtgattg
     5641 tgtttgtgtg tgtgattgtg tatatgtgtg tatggttgtg tgtgattgtg tgtatgtatg
     5701 tttgtgtgtg attgtgtgtg tgtgattgtg catgtgtgtg tgtgtgattg tgtttatgtg
     5761 tatgattgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgttgtgtat
     5821 atatatttat ggtagtgaga ggcaacgctc cggctcaggt gtcaggttgg tttttgagac
     5881 agagtctttc acttagcttg gaattaattc actggccgtc gttttacaac gtcgtgactg
     5941 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     6001 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     6061 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     6121 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     6181 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     6241 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     6301 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     6361 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     6421 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     6481 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     6541 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6601 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6661 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6721 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6781 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6841 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6901 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6961 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     7021 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     7081 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     7141 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     7201 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     7261 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     7321 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     7381 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     7441 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     7501 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     7561 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     7621 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7681 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7741 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7801 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7861 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7921 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7981 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     8041 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     8101 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     8161 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     8221 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     8281 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     8341 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     8401 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     8461 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     8521 ctatgaccat gattacgc
//
