LOCUS       sequence-246304-        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"
     CDS             14..55
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             62..1018
                     /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             1025..1051
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             join(1082..1084,1085..1087,1088..1801)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1388..1407)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1735..1756
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1824..1923)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1940..1959)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1941..1959)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1964..1980)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1964..1980)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2082..2101)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2093..2902
                     /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(2160..2179)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3049..3637
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3538..3557
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3791..3808
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3967..3994)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4086..4172)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4221..4243
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4235..4252
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4236..4252
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4268..4367
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ccgcggccgc atgggcaagc ccatccccaa ccccctgctg ggcctggaca gcaccgctag
       61 caaagacaat actgtgcctc tgaagctgat cgctctcctg gctaatggcg agttccatag
      121 tggcgaacag ctgggagaaa ccctgggcat gtccagggcc gctatcaaca agcacattca
      181 gactctgcgc gactggggcg tggacgtgtt caccgtgccc ggaaagggct actctctgcc
      241 cgagcctatc ccgctgctga acgctaaaca gattctggga cagctggacg gcgggagcgt
      301 ggcagtcctg cctgtggtcg actccaccaa tcagtacctg ctggatcgaa tcggcgagct
      361 gaagagtggg gatgcttgca ttgcagaata tcagcaggca gggagaggaa gcagagggag
      421 gaaatggttc tctccttttg gagctaacct gtacctgagt atgttttggc gcctgaagcg
      481 gggaccagca gcaatcggcc tgggcccggt catcggaatt gtcatggcag aagcgctgcg
      541 aaagctggga gcagacaagg tgcgagtcaa atggcccaat gacctgtatc tgcaggatag
      601 aaagctggca ggcatcctgg tggagctggc cggaataaca ggcgatgctg cacagatcgt
      661 cattggcgcc gggattaacg tggctatgag gcgcgtggag gaaagcgtgg tcaatcaggg
      721 ctggatcaca ctgcaggaag cagggattaa cctggacagg aatactctgg ccgctacgct
      781 gatccgagag ctgcgggcag ccctggaact gttcgagcag gaaggcctgg ctccatatct
      841 gccacggtgg gagaagctgg ataacttcat caatagaccc gtgaagctga tcattgggga
      901 caaagagatt ttcgggatta gccgggggat tgataaacag ggagccctgc tgctggaaca
      961 ggacggagtt atcaaaccct ggatgggcgg agaaatcagt ctgcggtctg ccgaaaagct
     1021 gcagctgcct cccctggagc gcctgaccct ggacggaggc ggtggaagcg gcggaggttc
     1081 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1141 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1201 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     1261 cctcgtgacc accttcggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa
     1321 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1381 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1441 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1501 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     1561 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     1621 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1681 ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     1741 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     1801 agggcccaag ggtgggcgcg ccgacccagc tttcttgtac aaagttggca ttataagaaa
     1861 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     1921 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     1981 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2041 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2101 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2161 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     2221 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2281 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     2341 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     2401 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     2461 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     2521 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     2581 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     2641 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     2701 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     2761 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     2821 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     2881 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     2941 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     3001 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3061 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3121 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3181 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3241 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3301 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3361 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3421 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3481 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3541 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3601 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3661 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3721 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3781 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     3841 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     3901 agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga
     3961 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat
     4021 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc
     4081 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
     4141 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt
     4201 aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct
     4261 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
     4321 tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gcaggct
//
