LOCUS Exported 4169 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION gateway entry vector for expressing nuclear miniTurbo-YFP under a promoter of choice. ACCESSION . VERSION . KEYWORDS pENTR_L1-miniTurbo-YFP-NLS-STOP-L2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4169) 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 4169) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4169 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind complement(17..116) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(133..152) /label=T7 /note="T7 promoter, forward primer" promoter complement(134..152) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(157..173) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(157..173) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(275..294) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 286..1095 /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(353..372) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 1242..1830 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1731..1750 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1984..2001 /label=L4440 /note="L4440 vector, forward primer" terminator 2160..2187 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 2279..2365 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 2414..2436 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2428..2445 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2429..2445 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 2461..2560 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 2574..2615 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 2622..3392 /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 3420..4136 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3465..3486) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3726..3745) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4073..4094 /label=EGFP-C /note="EGFP, forward primer" CDS 4146..4166 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" ORIGIN 1 aagggtgggc gcgccgaccc agctttcttg tacaaagttg gcattataag aaagcattgc 61 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat 121 ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc 181 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga 241 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa 301 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa 361 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc 421 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat 481 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt 541 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc 601 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc 661 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt 721 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat 781 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca 841 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac 901 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag 961 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt 1021 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc 1081 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg 1141 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc 1201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 1261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 1321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 1381 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 1441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 1501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 1561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 1621 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 1681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 1741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 1801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 1861 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg 1921 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag 1981 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc 2041 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc 2101 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa 2161 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc 2221 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat 2281 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt 2341 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta 2401 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc 2461 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa 2521 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgcggccgc atgggcaagc 2581 ccatccccaa ccccctgctg ggcctggaca gcaccgctag catcccgctg ctgaacgcta 2641 aacagattct gggacagctg gacggcggga gcgtggcagt cctgcctgtg gtcgactcca 2701 ccaatcagta cctgctggat cgaatcggcg agctgaagag tggggatgct tgcattgcag 2761 aatatcagca ggcagggaga ggaagcagag ggaggaaatg gttctctcct tttggagcta 2821 acctgtacct gagtatgttt tggcgcctga agcggggacc agcagcaatc ggcctgggcc 2881 cggtcatcgg aattgtcatg gcagaagcgc tgcgaaagct gggagcagac aaggtgcgag 2941 tcaaatggcc caatgacctg tatctgcagg atagaaagct ggcaggcatc ctggtggagc 3001 tggccggaat aacaggcgat gctgcacaga tcgtcattgg cgccgggatt aacgtggcta 3061 tgaggcgcgt ggaggaaagc gtggtcaatc agggctggat cacactgcag gaagcaggga 3121 ttaacctgga caggaatact ctggccgcta tgctgatccg agagctgcgg gcagccctgg 3181 aactgttcga gcaggaaggc ctggctccat atctgtcacg gtgggagaag ctggataact 3241 tcatcaatag acccgtgaag ctgatcattg gggacaaaga gattttcggg attagccggg 3301 ggattgataa acagggagcc ctgctgctgg aacaggacgg agttatcaaa ccctggatgg 3361 gcggagaaat cagtctgcgg tctgccgaaa agggaggcgg tggaagcggc ggaggttcca 3421 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg 3481 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg 3541 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc 3601 tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc 3661 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct 3721 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg 3781 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca 3841 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg 3901 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg 3961 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact 4021 acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc 4081 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaaggctt 4141 cggatccaaa aaagaagaga aaggtatag //