LOCUS Exported 4181 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION gateway entry vector for expressing cytosolic miniTurbo-YFP under a promoter of choice. ACCESSION . VERSION . KEYWORDS pENTR_L1-miniTurbo-NES-YFP-STOP-L2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4181) AUTHORS Mair A, Xu SL, Branon TC, Ting AY, Bergmann DC TITLE Proximity labeling of protein complexes and cell type-specific organellar proteomes in Arabidopsis enabled by TurboID. JOURNAL Elife. 2019 Sep 19;8. pii: 47864. doi: 10.7554/eLife.47864. PUBMED 31535972 REFERENCE 2 (bases 1 to 4181) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4181 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind complement(26..125) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(142..161) /label=T7 /note="T7 promoter, forward primer" promoter complement(143..161) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(166..182) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(166..182) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(284..303) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 295..1104 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(362..381) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 1251..1839 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1740..1759 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1993..2010 /label=L4440 /note="L4440 vector, forward primer" terminator 2169..2196 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 2288..2374 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 2423..2445 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2437..2454 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2438..2454 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 2470..2569 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 2583..2624 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 2631..3401 /codon_start=1 /product="N-terminally truncated mutant of the E. coli biotin ligase BioID engineered for improved catalytic efficiency (Branon et al., 2018)" /label=miniTurbo /translation="IPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGDACI AEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLGADK VRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWITLQ EAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIF GISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK" CDS 3408..3434 /codon_start=1 /product="nuclear export signal from the HIV Rev protein (Fischer et al., 1995)" /label=NES /translation="LPPLERLTL" CDS 3465..3 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3465..4181 /codon_start=1 /product="enhanced variant of YFP (Heikal et al., 2001)" /label=Citrine /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3510..3531) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3771..3790) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4118..4139 /label=EGFP-C /note="EGFP, forward primer" ORIGIN 1 taagggccca agggtgggcg cgccgaccca gctttcttgt acaaagttgg cattataaga 61 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt 121 atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc 181 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata 241 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc 301 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat 361 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat 421 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat 481 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc 541 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa 601 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg 661 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat 721 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt 781 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa 841 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt 901 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac 961 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag 1021 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat 1081 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc 1141 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac 1201 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 1261 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 1321 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 1381 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct 1441 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 1501 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 1561 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 1621 ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg 1681 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 1741 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 1801 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 1861 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 1921 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga 1981 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg 2041 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg 2101 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta 2161 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt 2221 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc 2281 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg 2341 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg 2401 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag 2461 ctcgggcccc aaataatgat tttattttga ctgatagtga cctgttcgtt gcaacaaatt 2521 gatgagcaat gcttttttat aatgccaact ttgtacaaaa aagcaggctc cgcggccgca 2581 tgggcaagcc catccccaac cccctgctgg gcctggacag caccgctagc atcccgctgc 2641 tgaacgctaa acagattctg ggacagctgg acggcgggag cgtggcagtc ctgcctgtgg 2701 tcgactccac caatcagtac ctgctggatc gaatcggcga gctgaagagt ggggatgctt 2761 gcattgcaga atatcagcag gcagggagag gaagcagagg gaggaaatgg ttctctcctt 2821 ttggagctaa cctgtacctg agtatgtttt ggcgcctgaa gcggggacca gcagcaatcg 2881 gcctgggccc ggtcatcgga attgtcatgg cagaagcgct gcgaaagctg ggagcagaca 2941 aggtgcgagt caaatggccc aatgacctgt atctgcagga tagaaagctg gcaggcatcc 3001 tggtggagct ggccggaata acaggcgatg ctgcacagat cgtcattggc gccgggatta 3061 acgtggctat gaggcgcgtg gaggaaagcg tggtcaatca gggctggatc acactgcagg 3121 aagcagggat taacctggac aggaatactc tggccgctat gctgatccga gagctgcggg 3181 cagccctgga actgttcgag caggaaggcc tggctccata tctgtcacgg tgggagaagc 3241 tggataactt catcaataga cccgtgaagc tgatcattgg ggacaaagag attttcggga 3301 ttagccgggg gattgataaa cagggagccc tgctgctgga acaggacgga gttatcaaac 3361 cctggatggg cggagaaatc agtctgcggt ctgccgaaaa gctgcagctg cctcccctgg 3421 agcgcctgac cctggacgga ggcggtggaa gcggcggagg ttccatggtg agcaagggcg 3481 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc 3541 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga 3601 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg 3661 gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca 3721 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca 3781 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc 3841 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact 3901 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact 3961 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga 4021 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt 4081 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga 4141 ccgccgccgg gatcactctc ggcatggacg agctgtacaa g //