LOCUS Exported 4179 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION gateway entry vector for making N-terminal YFP-miniTurbo fusions with nuclear and non-nuclear proteins. ACCESSION . VERSION . KEYWORDS pENTR_L1-YFP-miniTurbo-L2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4179) 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 4179) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4179 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..17 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 33..132 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 153..869 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(198..219) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(459..478) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 806..827 /label=EGFP-C /note="EGFP, forward primer" CDS 915..956 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 963..1733 /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" protein_bind complement(1768..1867) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1884..1903) /label=T7 /note="T7 promoter, forward primer" promoter complement(1885..1903) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1908..1924) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1908..1924) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2026..2045) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 2037..2846 /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(2104..2123) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2993..3581 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3482..3501 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3735..3752 /label=L4440 /note="L4440 vector, forward primer" terminator 3911..3938 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 4030..4116 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 4165..8 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4179..17 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" ORIGIN 1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag 61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca 121 aaaaagcagg ctccgcggcc gcccccttca ccatggtgag caagggcgag gagctgttca 181 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg 241 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca 301 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcctgc 361 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc 421 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc 481 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg 541 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca 601 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc 661 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg 721 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca 781 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga 841 tcactctcgg catggacgag ctgtacaagg ctaagggtgg gcgcgccggt ggaggcggtt 901 caggaggtgg catgggcaag cccatcccca accccctgct gggcctggac agcaccgcta 961 gcatcccgct gctgaacgct aaacagattc tgggacagct ggacggcggg agcgtggcag 1021 tcctgcctgt ggtcgactcc accaatcagt acctgctgga tcgaatcggc gagctgaaga 1081 gtggggatgc ttgcattgca gaatatcagc aggcagggag aggaagcaga gggaggaaat 1141 ggttctctcc ttttggagct aacctgtacc tgagtatgtt ttggcgcctg aagcggggac 1201 cagcagcaat cggcctgggc ccggtcatcg gaattgtcat ggcagaagcg ctgcgaaagc 1261 tgggagcaga caaggtgcga gtcaaatggc ccaatgacct gtatctgcag gatagaaagc 1321 tggcaggcat cctggtggag ctggccggaa taacaggcga tgctgcacag atcgtcattg 1381 gcgccgggat taacgtggct atgaggcgcg tggaggaaag cgtggtcaat cagggctgga 1441 tcacactgca ggaagcaggg attaacctgg acaggaatac tctggccgct atgctgatcc 1501 gagagctgcg ggcagccctg gaactgttcg agcaggaagg cctggctcca tatctgtcac 1561 ggtgggagaa gctggataac ttcatcaata gacccgtgaa gctgatcatt ggggacaaag 1621 agattttcgg gattagccgg gggattgata aacagggagc cctgctgctg gaacaggacg 1681 gagttatcaa accctggatg ggcggagaaa tcagtctgcg gtctgccgaa aagggaggcg 1741 gtggaagcgg cggaggtggg cgcgccgacc cagctttctt gtacaaagtt ggcattataa 1801 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca 1861 ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt 1921 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata 1981 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga 2041 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat 2101 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt 2161 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg 2221 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca 2281 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa 2341 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc 2401 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg 2461 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga 2521 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata 2581 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc 2641 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag 2701 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac 2761 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc 2821 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga 2881 gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt 2941 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc 3001 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 3061 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag 3121 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact 3181 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 3241 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 3301 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 3361 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 3421 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 3481 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc 3541 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct 3601 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc 3661 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc 3721 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac 3781 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact 3841 ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg 3901 tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt 3961 ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc 4021 tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa 4081 ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg 4141 cgttaacgct agcatggatg ttttcccagt cacgacgtt //