LOCUS       sequence-243660-        4367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway entry vector for expressing cytosolic TurboID-YFP under a 
            promoter of choice.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 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-NES-YFP-STOP-L2
FEATURES             Location/Qualifiers
     source          1..4367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(26..125)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(142..161)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(143..161)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(166..182)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(166..182)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(284..303)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             295..1104
                     /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(362..381)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1251..1839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1740..1759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1993..2010
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2169..2196)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2288..2374)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2423..2445
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2437..2454
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2438..2454
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2470..2569
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2583..2624
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2631..3587
                     /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             3594..3620
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             join(3651..3653,3654..3656,3657..4367,1..3)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3957..3976)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4304..4325
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 taagggccca agggtgggcg cgccgaccca gctttcttgt acaaagttgg cattataaga
       61 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
      121 atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
      181 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
      241 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
      301 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
      361 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
      421 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
      481 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
      541 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
      601 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
      661 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
      721 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
      781 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
      841 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
      901 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
      961 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     1021 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     1081 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     1141 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     1201 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1261 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1321 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1381 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     1441 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1501 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1561 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1621 ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1681 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1741 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1801 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1861 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     1921 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     1981 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     2041 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     2101 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
     2161 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     2221 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     2281 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     2341 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     2401 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
     2461 ctcgggcccc aaataatgat tttattttga ctgatagtga cctgttcgtt gcaacaaatt
     2521 gatgagcaat gcttttttat aatgccaact ttgtacaaaa aagcaggctc cgcggccgca
     2581 tgggcaagcc catccccaac cccctgctgg gcctggacag caccgctagc aaagacaata
     2641 ctgtgcctct gaagctgatc gctctcctgg ctaatggcga gttccatagt ggcgaacagc
     2701 tgggagaaac cctgggcatg tccagggccg ctatcaacaa gcacattcag actctgcgcg
     2761 actggggcgt ggacgtgttc accgtgcccg gaaagggcta ctctctgccc gagcctatcc
     2821 cgctgctgaa cgctaaacag attctgggac agctggacgg cgggagcgtg gcagtcctgc
     2881 ctgtggtcga ctccaccaat cagtacctgc tggatcgaat cggcgagctg aagagtgggg
     2941 atgcttgcat tgcagaatat cagcaggcag ggagaggaag cagagggagg aaatggttct
     3001 ctccttttgg agctaacctg tacctgagta tgttttggcg cctgaagcgg ggaccagcag
     3061 caatcggcct gggcccggtc atcggaattg tcatggcaga agcgctgcga aagctgggag
     3121 cagacaaggt gcgagtcaaa tggcccaatg acctgtatct gcaggataga aagctggcag
     3181 gcatcctggt ggagctggcc ggaataacag gcgatgctgc acagatcgtc attggcgccg
     3241 ggattaacgt ggctatgagg cgcgtggagg aaagcgtggt caatcagggc tggatcacac
     3301 tgcaggaagc agggattaac ctggacagga atactctggc cgctacgctg atccgagagc
     3361 tgcgggcagc cctggaactg ttcgagcagg aaggcctggc tccatatctg ccacggtggg
     3421 agaagctgga taacttcatc aatagacccg tgaagctgat cattggggac aaagagattt
     3481 tcgggattag ccgggggatt gataaacagg gagccctgct gctggaacag gacggagtta
     3541 tcaaaccctg gatgggcgga gaaatcagtc tgcggtctgc cgaaaagctg cagctgcctc
     3601 ccctggagcg cctgaccctg gacggaggcg gtggaagcgg cggaggttcc atggtgagca
     3661 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     3721 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     3781 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     3841 ccttcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     3901 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     3961 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     4021 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     4081 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     4141 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     4201 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     4261 accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     4321 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaag
//
