LOCUS       sequence-246303-        4365 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway entry vector for making N-terminal YFP-TurboID fusions with 
            nuclear and non-nuclear proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4365)
  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 4365)
  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-YFP-Turbo-L2
FEATURES             Location/Qualifiers
     source          1..4365
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(153..155,156..158,159..869)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     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..1919
                     /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"
     protein_bind    complement(1954..2053)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2070..2089)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2071..2089)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2094..2110)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2094..2110)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2212..2231)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2223..3032
                     /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(2290..2309)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3179..3767
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3668..3687
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3921..3938
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4097..4124)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4216..4302)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4351..4365,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4365,1..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 gcaaagacaa tactgtgcct ctgaagctga tcgctctcct ggctaatggc gagttccata
     1021 gtggcgaaca gctgggagaa accctgggca tgtccagggc cgctatcaac aagcacattc
     1081 agactctgcg cgactggggc gtggacgtgt tcaccgtgcc cggaaagggc tactctctgc
     1141 ccgagcctat cccgctgctg aacgctaaac agattctggg acagctggac ggcgggagcg
     1201 tggcagtcct gcctgtggtc gactccacca atcagtacct gctggatcga atcggcgagc
     1261 tgaagagtgg ggatgcttgc attgcagaat atcagcaggc agggagagga agcagaggga
     1321 ggaaatggtt ctctcctttt ggagctaacc tgtacctgag tatgttttgg cgcctgaagc
     1381 ggggaccagc agcaatcggc ctgggcccgg tcatcggaat tgtcatggca gaagcgctgc
     1441 gaaagctggg agcagacaag gtgcgagtca aatggcccaa tgacctgtat ctgcaggata
     1501 gaaagctggc aggcatcctg gtggagctgg ccggaataac aggcgatgct gcacagatcg
     1561 tcattggcgc cgggattaac gtggctatga ggcgcgtgga ggaaagcgtg gtcaatcagg
     1621 gctggatcac actgcaggaa gcagggatta acctggacag gaatactctg gccgctacgc
     1681 tgatccgaga gctgcgggca gccctggaac tgttcgagca ggaaggcctg gctccatatc
     1741 tgccacggtg ggagaagctg gataacttca tcaatagacc cgtgaagctg atcattgggg
     1801 acaaagagat tttcgggatt agccggggga ttgataaaca gggagccctg ctgctggaac
     1861 aggacggagt tatcaaaccc tggatgggcg gagaaatcag tctgcggtct gccgaaaagg
     1921 gaggcggtgg aagcggcgga ggtgggcgcg ccgacccagc tttcttgtac aaagttggca
     1981 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     2041 aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag
     2101 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     2161 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     2221 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     2281 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     2341 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     2401 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     2461 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     2521 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     2581 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     2641 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     2701 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     2761 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     2821 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     2881 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     2941 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     3001 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     3061 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     3121 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3181 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3241 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3301 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3361 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3421 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3481 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3541 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3601 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3661 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3721 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3781 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3841 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3901 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     3961 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     4021 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
     4081 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
     4141 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
     4201 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     4261 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
     4321 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgtt
//
