LOCUS       sequence-243661-        4355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway entry vector for expressing nuclear TurboID-YFP under a 
            promoter of choice.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4355)
  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 4355)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2019-09-19"; volume: "8. pii: 47864. doi"; pages: " 
            10.7554/eLife.47864"
COMMENT     Sequence Label: pENTR_L1-Turbo-YFP-NLS-STOP-L2
FEATURES             Location/Qualifiers
     source          1..4355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             10..51
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             58..1014
                     /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             join(1042..1044,1045..1047,1048..1758)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1348..1367)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1695..1716
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1768..1788
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(1808..1907)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1924..1943)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1925..1943)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1948..1964)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1948..1964)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2066..2085)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2077..2886
                     /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(2144..2163)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3033..3621
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3522..3541
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3775..3792
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3951..3978)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4070..4156)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4205..4227
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4219..4236
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4220..4236
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4252..4351
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ggccgcatgg gcaagcccat ccccaacccc ctgctgggcc tggacagcac cgctagcaaa
       61 gacaatactg tgcctctgaa gctgatcgct ctcctggcta atggcgagtt ccatagtggc
      121 gaacagctgg gagaaaccct gggcatgtcc agggccgcta tcaacaagca cattcagact
      181 ctgcgcgact ggggcgtgga cgtgttcacc gtgcccggaa agggctactc tctgcccgag
      241 cctatcccgc tgctgaacgc taaacagatt ctgggacagc tggacggcgg gagcgtggca
      301 gtcctgcctg tggtcgactc caccaatcag tacctgctgg atcgaatcgg cgagctgaag
      361 agtggggatg cttgcattgc agaatatcag caggcaggga gaggaagcag agggaggaaa
      421 tggttctctc cttttggagc taacctgtac ctgagtatgt tttggcgcct gaagcgggga
      481 ccagcagcaa tcggcctggg cccggtcatc ggaattgtca tggcagaagc gctgcgaaag
      541 ctgggagcag acaaggtgcg agtcaaatgg cccaatgacc tgtatctgca ggatagaaag
      601 ctggcaggca tcctggtgga gctggccgga ataacaggcg atgctgcaca gatcgtcatt
      661 ggcgccggga ttaacgtggc tatgaggcgc gtggaggaaa gcgtggtcaa tcagggctgg
      721 atcacactgc aggaagcagg gattaacctg gacaggaata ctctggccgc tacgctgatc
      781 cgagagctgc gggcagccct ggaactgttc gagcaggaag gcctggctcc atatctgcca
      841 cggtgggaga agctggataa cttcatcaat agacccgtga agctgatcat tggggacaaa
      901 gagattttcg ggattagccg ggggattgat aaacagggag ccctgctgct ggaacaggac
      961 ggagttatca aaccctggat gggcggagaa atcagtctgc ggtctgccga aaagggaggc
     1021 ggtggaagcg gcggaggttc catggtgagc aagggcgagg agctgttcac cggggtggtg
     1081 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1141 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1201 ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca gtgcttcgcc
     1261 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1321 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1381 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1441 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1501 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     1561 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1621 gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac
     1681 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1741 atggacgagc tgtacaaggc ttcggatcca aaaaagaaga gaaaggtata gaagggtggg
     1801 cgcgccgacc cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt
     1861 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat
     1921 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
     1981 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     2041 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     2101 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     2161 gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca
     2221 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     2281 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     2341 cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta
     2401 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     2461 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     2521 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     2581 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     2641 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     2701 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     2761 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     2821 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     2881 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     2941 gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     3001 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3061 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3121 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     3181 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3241 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3301 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3361 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3421 gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3481 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3541 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3601 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3661 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3721 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3781 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     3841 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     3901 aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat
     3961 ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc
     4021 gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac
     4081 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag
     4141 cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg
     4201 ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc ccaaataatg
     4261 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgatgagca atgctttttt
     4321 ataatgccaa ctttgtacaa aaaagcaggc tccgc
//
