LOCUS       sequence-246302-        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"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             114..155
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             162..1118
                     /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(1146..1148,1149..1151,1152..1862)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1452..1471)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1799..1820
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1872..1892
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(1912..2011)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2028..2047)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2029..2047)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2052..2068)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2052..2068)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2170..2189)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2181..2990
                     /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(2248..2267)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3137..3725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3626..3645
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3879..3896
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4055..4082)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4174..4260)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4309..4331
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4323..4340
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4324..4340
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgcggccgc atgggcaagc
      121 ccatccccaa ccccctgctg ggcctggaca gcaccgctag caaagacaat actgtgcctc
      181 tgaagctgat cgctctcctg gctaatggcg agttccatag tggcgaacag ctgggagaaa
      241 ccctgggcat gtccagggcc gctatcaaca agcacattca gactctgcgc gactggggcg
      301 tggacgtgtt caccgtgccc ggaaagggct actctctgcc cgagcctatc ccgctgctga
      361 acgctaaaca gattctggga cagctggacg gcgggagcgt ggcagtcctg cctgtggtcg
      421 actccaccaa tcagtacctg ctggatcgaa tcggcgagct gaagagtggg gatgcttgca
      481 ttgcagaata tcagcaggca gggagaggaa gcagagggag gaaatggttc tctccttttg
      541 gagctaacct gtacctgagt atgttttggc gcctgaagcg gggaccagca gcaatcggcc
      601 tgggcccggt catcggaatt gtcatggcag aagcgctgcg aaagctggga gcagacaagg
      661 tgcgagtcaa atggcccaat gacctgtatc tgcaggatag aaagctggca ggcatcctgg
      721 tggagctggc cggaataaca ggcgatgctg cacagatcgt cattggcgcc gggattaacg
      781 tggctatgag gcgcgtggag gaaagcgtgg tcaatcaggg ctggatcaca ctgcaggaag
      841 cagggattaa cctggacagg aatactctgg ccgctacgct gatccgagag ctgcgggcag
      901 ccctggaact gttcgagcag gaaggcctgg ctccatatct gccacggtgg gagaagctgg
      961 ataacttcat caatagaccc gtgaagctga tcattgggga caaagagatt ttcgggatta
     1021 gccgggggat tgataaacag ggagccctgc tgctggaaca ggacggagtt atcaaaccct
     1081 ggatgggcgg agaaatcagt ctgcggtctg ccgaaaaggg aggcggtgga agcggcggag
     1141 gttccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1201 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1261 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1321 ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
     1381 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1441 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1501 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1561 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1621 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1681 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1741 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1801 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1861 aggcttcgga tccaaaaaag aagagaaagg tatagaaggg tgggcgcgcc gacccagctt
     1921 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg
     1981 tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt
     2041 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat
     2101 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     2161 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa
     2221 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
     2281 caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac
     2341 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
     2401 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
     2461 tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt
     2521 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
     2581 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
     2641 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     2701 aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc
     2761 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     2821 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     2881 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     2941 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg
     3001 ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat
     3061 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
     3121 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3181 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3241 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3301 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3361 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3421 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3481 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3541 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3601 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3661 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3721 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3781 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3841 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     3901 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     3961 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc
     4021 gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt
     4081 ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg
     4141 ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac
     4201 cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg
     4261 atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac
     4321 gttgtaaaac gacggccagt cttaagctcg ggccc
//
