LOCUS pSF3-Flag-CBir-FRB_Myc-NBir-FKBP 8529 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Split-BioID plasmid: Expresses N-terminally tagged CBir[257-321]-FRB
and N-terminally tagged NBir[2-256]-FKBP.
ACCESSION .
VERSION .
KEYWORDS pSF3-Flag-CBir-FRB_Myc-NBir-FKBP
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 May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8529
/organism="synthetic DNA construct"
/mol_type="other DNA"
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"
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(2382..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 3715..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 6486..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 8429..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
//
