LOCUS Exported 6411 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION To fuse your protein of interest to the C-terminus of BirA(R118G) and use in proximity-dependent biotin identification (BioID); myc tag. ACCESSION . VERSION . KEYWORDS pcDNA3.1 mycBioID SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6411) AUTHORS Roux KJ, Kim DI, Raida M, Burke B TITLE A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins in mammalian cells. JOURNAL J Cell Biol. 2012 Mar 12. PUBMED 22412018 REFERENCE 2 (bases 1 to 6411) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6411 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 899..908 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 908..937 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 941..1900 /codon_start=1 /product="promiscuous R118G mutant of the E. coli biotin protein ligase (Cronan, 2005)" /label=BirA(R118G) /note="also known as BirA* or BioID" /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK" primer_bind complement(2001..2018) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2007..2231 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2277..2705 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2364..2383) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2574..2595 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2714..2734) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2719..3049 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2900..3035 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2962..2981 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3116..3910 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3170..3189) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3780..3799 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4084..4205 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4121..4140) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4175..4194 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4254..4270) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4254..4270) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4267..4289) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4278..4294 /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(4302..4332) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4347..4368 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4485..4502) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4656..5244) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4736..4755) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5415..6275) /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 6038..6057 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6276..6380) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 caccatggaa caaaaactca tctcagaaga ggatctcgac aaggacaaca ccgtgcccct 961 gaagctgatc gccctgctgg ccaacggcga gttccactct ggcgagcagc tgggagagac 1021 cctgggaatg agcagagccg ccatcaacaa gcacatccag acactgagag actggggagt 1081 ggacgtgttc accgtgcctg gcaagggcta cagcctgcct gagcctatcc agctgctgaa 1141 cgccaagcag atcctgggac agctggatgg cggaagcgtg gccgtgctgc ctgtgatcga 1201 ctccaccaat cagtacctgc tggacagaat cggagagctg aagtccggcg acgcctgcat 1261 cgccgagtac cagcaggctg gcagaggagg cagaggacgg aagtggttca gcccattcgg 1321 agccaacctg tacctgtcca tgttctggag actggagcag ggacctgctg ctgccatcgg 1381 actgagtctg gtgatcggaa tcgtgatggc cgaggtgctg agaaagctgg gagccgacaa 1441 ggtgagagtg aagtggccta atgacctgta cctccaggac cgcaagctgg ctggcatcct 1501 ggtggagctg acaggcaaga caggcgatgc cgctcagatc gtgatcggag ccggaatcaa 1561 catggccatg agaagagtgg aggagagcgt ggtgaaccag ggctggatca ccctgcagga 1621 ggctggcatc aacctggacc ggaacaccct ggccgccatg ctgatcagag agctgagagc 1681 cgctctggag ctgttcgagc aggagggact ggctccttac ctgagcagat gggagaagct 1741 ggacaacttc atcaacagac ctgtgaagct gatcatcggc gacaaggaaa tcttcggcat 1801 ctccagagga atcgacaagc agggagctct gctgctggag caggacggaa tcatcaagcc 1861 ctggatgggc ggagaaatct ccctgagaag cgcagagaag ctcgagcggc cgccactgtg 1921 ctggatatct gcagaattcc accacactgg actagtggat ccgagctcgg taccaagctt 1981 aagtttaaac cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 2041 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 2101 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 2161 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt 2221 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc 2281 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 2341 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 2401 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc 2461 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 2521 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 2581 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 2641 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 2701 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccaggc 2761 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc 2821 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt 2881 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc 2941 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct 3001 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg 3061 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat 3121 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta 3181 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca 3241 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga 3301 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga 3361 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct 3421 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg 3481 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga 3541 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca 3601 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga 3661 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg 3721 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc 3781 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt 3841 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga 3901 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca 3961 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc 4021 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac 4081 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 4141 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 4201 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag 4261 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 4321 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 4381 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 4441 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 4501 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 4561 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 4621 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 4681 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 4741 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 4801 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc 4861 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 4921 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 4981 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 5041 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca 5101 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 5161 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 5221 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 5281 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 5341 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 5401 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 5461 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 5521 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 5581 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 5641 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 5701 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 5761 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 5821 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 5881 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 5941 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 6001 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 6061 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 6121 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 6181 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 6241 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 6301 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 6361 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c //