LOCUS       sequence-182094-        8529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Split-BioID plasmid: Expresses N-terminally tagged CBir[257-321]-FRB
            and N-terminally tagged NBir[2-256]-FKBP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8529)
  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 8529)
  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_Myc-NBir-FKBP
FEATURES             Location/Qualifiers
     source          1..8529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             508..977
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    1042..1399
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     primer_bind     1848..1864
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    1917..1950
                     /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    2058..2192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2095..2114)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(2382..2702,2703..2705))
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             complement(3534..3563)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(3661..3698)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    join(3715..3733,3734..3756,3757..3775,3776..3798,3799..3817,
                     3818..3840,3841..3859,3860..3882,3883..3901,3902..3924,
                     3925..3943,3944..3966,3967..3985)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        4000..4038
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             4150..4173
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4441..4719
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     CDS             4453..4725
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQL"
     protein_bind    4760..4807
                     /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(5891..5908)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5891..5907)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5900..5922)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6116..6135)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6235..6257
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6295..6313)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6381..6485
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6486..6554,6555..7346)
                     /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(6704..6723)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7517..8105
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8006..8025
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8259..8276
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8393..8414
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8429..8434,8435..8452,8453..8459)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8467..8483
                     /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     8472..8494
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8491..8507
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8491..8507
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tgctcttagg agtttcctaa tacatcccaa actcaaatat ataaagcatt tgacttgttc
       61 tatgccctag ggggcggggg gaagctaagc cagctttttt taacatttaa aatgttaatt
      121 ccattttaaa tgcacagatg tttttatttc ataagggttt caatgtgcat gaatgctgca
      181 atattcctgt taccaaagct agtataaata aaaatagata aacgtggaaa ttacttagag
      241 tttctgtcat taacgtttcc ttcctcagtt gacaacataa atgcgctgct gagaagccag
      301 tttgcatctg tcaggatcaa tttcccatta tgccagtcat attaattact agtcaattag
      361 ttgattttta tttttgacat atacatgtga aagaccccac ctgtaggttt ggcaagctag
      421 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gaaaagttca
      481 gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
      541 gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca
      601 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
      661 tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
      721 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
      781 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca
      841 gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
      901 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
      961 tcagcggggg tctttcattt gggggctcgt ccgggatcgg gagacccctg cccagggacc
     1021 accgacccac caccgggagg taagctggcc agcaacttat ctgtgtctgt ccgattgtct
     1081 agtgtctatg actgatttta tgcgcctgcg tcggtactag ttagctaact agctctgtat
     1141 ctggcggacc cgtggtggaa ctgacgagtt cggaacaccc ggccgcaacc ctgggagacg
     1201 tcccagggac ttcgggggcc gtttttgtgg cccgacctga gtcctaaaat cccgatcgtt
     1261 taggactctt tggtgcaccc cccttagagg agggatatgt ggttctggta ggagacgaga
     1321 acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttgggac cgaagccgcg
     1381 ccgcgcgtct tgtctgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt
     1441 atttgtctga aaatatgggc ccgggctagc ctgttaccac tcccttaagt ttgaccttag
     1501 gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac
     1561 gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg
     1621 gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc
     1681 atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc
     1741 ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc
     1801 cgtctctccc ccttgaacca cctcgttcga ccccgcctcg atcctccctt tatccagccc
     1861 tcactccttc tctaggcgcc cccatatggc catatgagat ccgaagttcc tattccgaag
     1921 ttcctattct tcaaatagta taggaacttc agatcataat cagccatacc acatttgtag
     1981 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2041 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaggcaata
     2101 gcatcacaaa tttcacaaat aaggcatttt tttcactgca ttctagtttt ggtttgtcca
     2161 aactcatcaa tgtatcttat catgtctgga tctcaaatcc ctcggaagct gcgcctgtct
     2221 taggttggag tgatacattt ttatcacttt tacccgtctt tggattaggc agtagctctg
     2281 acggccctcc tgtcttaggt tagtgaaaaa tgtcactctc ttacccgtca ttggctgtcc
     2341 agcttagctc gcaggggagg tggtctggat ccctaacgcg tttccagttt tagaagctcc
     2401 acatcgaaga cgagagtggc atgtggtggg atgatgcctg ggtgcccagt ggcaccatag
     2461 gcataatctg gagatatagt cagtttggct ctctgaccca cactcatctg ggcaacccct
     2521 tcttcccagc ctcggatcac ctcctgcttg cctagcataa acttaaaggg cttgtttctg
     2581 tcccgggagg aatcaaattt ctttccatct tcaagcatcc cggtgtagtg caccacgcag
     2641 gtctggccgc gcttggggaa ggtgcgcccg tctcctgggg agatggtttc cacctgcact
     2701 cccatatcga tgccccggga ggcgtaggag atctggccgc cgccggagga gccgccgcca
     2761 tgtttctcct gctcgaacag ctccagagcg gctctcagct ctctgatcag catggcggcc
     2821 agggtgttcc ggtccaggtt gatgccagcc tcctgcaggg tgatccagcc ctggttcacc
     2881 acgctctcct ccactcttct catggccatg ttgattccgg ctccgatcac gatctgagcg
     2941 gcatcgcctg tcttgcctgt cagctccacc aggatgccag ccagcttgcg gtcctggagg
     3001 tacaggtcat taggccactt cactctcacc ttgtcggctc ccagctttct cagcacctcg
     3061 gccatcacga ttccgatcac cagactcagt ccgatggcag cagcaggtcc ctgctccagt
     3121 ctccagaaca tggacaggta caggttggct ccgaatgggc tgaaccactt ccgtcctctg
     3181 cctcctctgc cagcctgctg gtactcggcg atgcaggcgt cgccggactt cagctctccg
     3241 attctgtcca gcaggtactg attggtggag tcgatcacag gcagcacggc cacgcttccg
     3301 ccatccagct gtcccaggat ctgcttggcg ttcagcagct ggataggctc aggcaggctg
     3361 tagcccttgc caggcacggt gaacacgtcc actccccagt ctctcagtgt ctggatgtgc
     3421 ttgttgatgg cggctctgct cattcccagg gtctctccca gctgctcgcc agagtggaac
     3481 tcgccgttgg ccagcagggc gatcagcttc aggggcacgg tgttgtcctt gtcgagatcc
     3541 tcttctgaga tgagtttttg ttccatctcg accgcggagg ctggatcggt cccggtgtct
     3601 tctatggagg tcaaaacagc gtggatggcg tctccaggcg atctgacggt tcactaaacg
     3661 agctctgctt atataggcct cccaccgtac acgcctacaa gcttctttca cttttctcta
     3721 tcactgatag ggagtggtaa actcgacttt cacttttctc tatcactgat agggagtggt
     3781 aaactcgact ttcacttttc tctatcactg atagggagtg gtaaactcga ctttcacttt
     3841 tctctatcac tgatagggag tggtaaactc gactttcact tttctctatc actgataggg
     3901 agtggtaaac tcgactttca cttttctcta tcactgatag ggagtggtaa actcgacttt
     3961 cacttttctc tatcactgat agggagtggt aaactcgagg gtaggcgtgt acggtgggag
     4021 gcctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
     4081 tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggaat tcggccggcc
     4141 tcaattatgg actacaagga cgacgatgac aagattaacg gactggctcc ttacctgagc
     4201 agatgggaga agctggacaa cttcatcaac agacctgtga agctgatcat cggcgacaag
     4261 gaaatcttcg gcatctccag aggaatcgac aagcagggag ctctgctgct ggagcaggac
     4321 ggaatcatca agccctggat gggcggagaa atctccctga gaagcgcaga gaagctcgag
     4381 gcgaaccaga tctcctacgc ctcccggggc ggcggctcct ccggcggcgg cgtttacatg
     4441 atcctctggc atgagatgtg gcatgaaggc ctggaagagg catctcgttt gtactttggg
     4501 gaaaggaacg tgaaaggcat gtttgaggtg ctggagccct tgcatgctat gatggaacgg
     4561 ggcccccaga ctctgaagga aacatccttt aatcaggcct atggtcgaga tttaatggag
     4621 gcccaagagt ggtgcaggaa gtacatgaaa tcagggaatg tcaaggacct cacccaagcc
     4681 tgggacctct attatcatgt gttccgacga atctcaaagc agttaattaa atagcgatta
     4741 ataacgagga tctggccgcg aagttcctat tccgaagttc ctattctcta gaaagtatag
     4801 gaacttcaga tccggattag tccaatttgt taaagacagg atatcagtgg tccaggctct
     4861 agttttgact caacaatatc accagctgaa gcctatagag tacgagccat agataaaata
     4921 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     4981 caagctagct cgacttctgt tcgcgcgctt ctgctccccg agctcgagcc cacaacccct
     5041 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     5101 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     5161 tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa ttaatttggt
     5221 tttttttctt aagtatttac attaaatggc catagtactt aaagttacat tggcttcctt
     5281 gaaataaaca tggagtattc agaatgtgtc ataaatattt ctaattttaa gatagtatct
     5341 ccattggctt tctacttttt cttttatttt tttttgtcct ctgtcttcca tttgttgttg
     5401 ttgttgtttg tttgtttgtt tgttggttgg ttggttaatt tttttttaaa gatcctacac
     5461 tatagttcaa gctagactat tagctactct gtaacccagg gtgaccttga agtcatgggt
     5521 agcctgctgt tttagccttc ccacatctaa gattacaggt atgagctatc atttttggta
     5581 tattgattga ttgattgatt gatgtgtgtg tgtgtgattg tgtttgtgtg tgtgattgtg
     5641 tatatgtgtg tatggttgtg tgtgattgtg tgtatgtatg tttgtgtgtg attgtgtgtg
     5701 tgtgattgtg catgtgtgtg tgtgtgattg tgtttatgtg tatgattgtg tgtgtgtgtg
     5761 tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgttgtgtat atatatttat ggtagtgaga
     5821 ggcaacgctc cggctcaggt gtcaggttgg tttttgagac agagtctttc acttagcttg
     5881 gaattaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     5941 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     6001 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     6061 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     6121 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     6181 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     6241 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     6301 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     6361 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6421 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6481 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6541 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6601 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6661 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6721 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6781 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6841 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6901 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6961 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     7021 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     7081 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     7141 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     7201 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     7261 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     7321 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     7381 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7441 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7501 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7561 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7621 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     7681 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7741 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7801 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7861 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7921 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7981 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     8041 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     8101 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     8161 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     8221 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     8281 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     8341 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     8401 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     8461 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     8521 agctagctt
//
