LOCUS       sequence-243651-        4358 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 4358)
  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 4358)
  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..4358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             760..801
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             808..1764
                     /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(1799..1898)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1915..1934)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1916..1934)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1939..1955)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1939..1955)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2057..2076)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2068..2877
                     /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(2135..2154)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3024..3612
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3513..3532
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3766..3783
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3942..3969)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4061..4147)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4196..4218
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4210..4227
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4211..4227
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4243..4342
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      481 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagaag
      721 ggtgggcgcg ccggtggagg cggttcagga ggtggcatgg gcaagcccat ccccaacccc
      781 ctgctgggcc tggacagcac cgctagcaaa gacaatactg tgcctctgaa gctgatcgct
      841 ctcctggcta atggcgagtt ccatagtggc gaacagctgg gagaaaccct gggcatgtcc
      901 agggccgcta tcaacaagca cattcagact ctgcgcgact ggggcgtgga cgtgttcacc
      961 gtgcccggaa agggctactc tctgcccgag cctatcccgc tgctgaacgc taaacagatt
     1021 ctgggacagc tggacggcgg gagcgtggca gtcctgcctg tggtcgactc caccaatcag
     1081 tacctgctgg atcgaatcgg cgagctgaag agtggggatg cttgcattgc agaatatcag
     1141 caggcaggga gaggaagcag agggaggaaa tggttctctc cttttggagc taacctgtac
     1201 ctgagtatgt tttggcgcct gaagcgggga ccagcagcaa tcggcctggg cccggtcatc
     1261 ggaattgtca tggcagaagc gctgcgaaag ctgggagcag acaaggtgcg agtcaaatgg
     1321 cccaatgacc tgtatctgca ggatagaaag ctggcaggca tcctggtgga gctggccgga
     1381 ataacaggcg atgctgcaca gatcgtcatt ggcgccggga ttaacgtggc tatgaggcgc
     1441 gtggaggaaa gcgtggtcaa tcagggctgg atcacactgc aggaagcagg gattaacctg
     1501 gacaggaata ctctggccgc tacgctgatc cgagagctgc gggcagccct ggaactgttc
     1561 gagcaggaag gcctggctcc atatctgcca cggtgggaga agctggataa cttcatcaat
     1621 agacccgtga agctgatcat tggggacaaa gagattttcg ggattagccg ggggattgat
     1681 aaacagggag ccctgctgct ggaacaggac ggagttatca aaccctggat gggcggagaa
     1741 atcagtctgc ggtctgccga aaagggaggc ggtggaagcg gcggaggtgg gcgcgccgac
     1801 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     1861 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat
     1921 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     1981 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     2041 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     2101 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     2161 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     2221 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     2281 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     2341 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     2401 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     2461 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     2521 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     2581 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     2641 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     2701 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     2761 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     2821 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     2881 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     2941 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     3001 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3061 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3121 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3181 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3241 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3301 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3361 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga
     3421 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3481 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3541 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3601 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3661 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3721 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3781 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     3841 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     3901 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     3961 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     4021 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     4081 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     4141 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag
     4201 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt
     4261 tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca
     4321 actttgtaca aaaaagcagg ctccgcggcc gccccctt
//
