LOCUS Exported 4367 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION gateway entry vector for expressing cytosolic TurboID-YFP under a promoter of choice. ACCESSION . VERSION . KEYWORDS pENTR_L1-Turbo-NES-YFP-STOP-L2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4367) 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 4367) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4367 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 14..55 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 62..1018 /codon_start=1 /product="mutant of the E. coli biotin ligase BioID engineered for improved catalytic efficiency (Branon et al., 2018)" /label=TurboID /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK" CDS 1025..1051 /codon_start=1 /product="nuclear export signal from the HIV Rev protein (Fischer et al., 1995)" /label=NES /translation="LPPLERLTL" CDS 1082..1801 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1127..1148) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1388..1407) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1735..1756 /label=EGFP-C /note="EGFP, forward primer" protein_bind complement(1824..1923) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1940..1959) /label=T7 /note="T7 promoter, forward primer" promoter complement(1941..1959) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1964..1980) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1964..1980) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2082..2101) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 2093..2902 /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(2160..2179) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 3049..3637 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3538..3557 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3791..3808 /label=L4440 /note="L4440 vector, forward primer" terminator 3967..3994 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 4086..4172 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 4221..4243 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4235..4252 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 4236..4252 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 4268..4367 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" ORIGIN 1 ccgcggccgc atgggcaagc ccatccccaa ccccctgctg ggcctggaca gcaccgctag 61 caaagacaat actgtgcctc tgaagctgat cgctctcctg gctaatggcg agttccatag 121 tggcgaacag ctgggagaaa ccctgggcat gtccagggcc gctatcaaca agcacattca 181 gactctgcgc gactggggcg tggacgtgtt caccgtgccc ggaaagggct actctctgcc 241 cgagcctatc ccgctgctga acgctaaaca gattctggga cagctggacg gcgggagcgt 301 ggcagtcctg cctgtggtcg actccaccaa tcagtacctg ctggatcgaa tcggcgagct 361 gaagagtggg gatgcttgca ttgcagaata tcagcaggca gggagaggaa gcagagggag 421 gaaatggttc tctccttttg gagctaacct gtacctgagt atgttttggc gcctgaagcg 481 gggaccagca gcaatcggcc tgggcccggt catcggaatt gtcatggcag aagcgctgcg 541 aaagctggga gcagacaagg tgcgagtcaa atggcccaat gacctgtatc tgcaggatag 601 aaagctggca ggcatcctgg tggagctggc cggaataaca ggcgatgctg cacagatcgt 661 cattggcgcc gggattaacg tggctatgag gcgcgtggag gaaagcgtgg tcaatcaggg 721 ctggatcaca ctgcaggaag cagggattaa cctggacagg aatactctgg ccgctacgct 781 gatccgagag ctgcgggcag ccctggaact gttcgagcag gaaggcctgg ctccatatct 841 gccacggtgg gagaagctgg ataacttcat caatagaccc gtgaagctga tcattgggga 901 caaagagatt ttcgggatta gccgggggat tgataaacag ggagccctgc tgctggaaca 961 ggacggagtt atcaaaccct ggatgggcgg agaaatcagt ctgcggtctg ccgaaaagct 1021 gcagctgcct cccctggagc gcctgaccct ggacggaggc ggtggaagcg gcggaggttc 1081 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga 1141 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta 1201 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac 1261 cctcgtgacc accttcggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa 1321 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt 1381 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct 1441 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca 1501 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa 1561 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc 1621 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca 1681 ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt 1741 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta 1801 agggcccaag ggtgggcgcg ccgacccagc tttcttgtac aaagttggca ttataagaaa 1861 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat 1921 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg 1981 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc 2041 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca 2101 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg 2161 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg 2221 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt 2281 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa 2341 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac 2401 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc 2461 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg 2521 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga 2581 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact 2641 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat 2701 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg 2761 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa 2821 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt 2881 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat 2941 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc 3001 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 3061 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 3121 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 3181 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 3241 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 3301 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 3361 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 3421 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 3481 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 3541 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 3601 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 3661 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga 3721 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac 3781 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc 3841 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa 3901 agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga 3961 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat 4021 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc 4081 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc 4141 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt 4201 aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct 4261 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga 4321 tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gcaggct //