LOCUS       sequence-246314-        4179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway entry vector for making N-terminal YFP-miniTurbo fusions 
            with nuclear and non-nuclear proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4179)
  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 4179)
  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-miniTurbo-L2
FEATURES             Location/Qualifiers
     source          1..4179
                     /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..1733
                     /codon_start=1
                     /product="N-terminally truncated mutant of the E. coli 
                     biotin ligase BioID engineered for improved catalytic 
                     efficiency (Branon et al., 2018)"
                     /label=miniTurbo
                     /translation="IPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGDACI
                     AEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLGADK
                     VRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWITLQ
                     EAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIF
                     GISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     protein_bind    complement(1768..1867)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1884..1903)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1885..1903)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1908..1924)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1908..1924)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2026..2045)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2037..2846
                     /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(2104..2123)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2993..3581
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3482..3501
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3735..3752
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3911..3938)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4030..4116)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4165..4179,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4179,1..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 gcatcccgct gctgaacgct aaacagattc tgggacagct ggacggcggg agcgtggcag
     1021 tcctgcctgt ggtcgactcc accaatcagt acctgctgga tcgaatcggc gagctgaaga
     1081 gtggggatgc ttgcattgca gaatatcagc aggcagggag aggaagcaga gggaggaaat
     1141 ggttctctcc ttttggagct aacctgtacc tgagtatgtt ttggcgcctg aagcggggac
     1201 cagcagcaat cggcctgggc ccggtcatcg gaattgtcat ggcagaagcg ctgcgaaagc
     1261 tgggagcaga caaggtgcga gtcaaatggc ccaatgacct gtatctgcag gatagaaagc
     1321 tggcaggcat cctggtggag ctggccggaa taacaggcga tgctgcacag atcgtcattg
     1381 gcgccgggat taacgtggct atgaggcgcg tggaggaaag cgtggtcaat cagggctgga
     1441 tcacactgca ggaagcaggg attaacctgg acaggaatac tctggccgct atgctgatcc
     1501 gagagctgcg ggcagccctg gaactgttcg agcaggaagg cctggctcca tatctgtcac
     1561 ggtgggagaa gctggataac ttcatcaata gacccgtgaa gctgatcatt ggggacaaag
     1621 agattttcgg gattagccgg gggattgata aacagggagc cctgctgctg gaacaggacg
     1681 gagttatcaa accctggatg ggcggagaaa tcagtctgcg gtctgccgaa aagggaggcg
     1741 gtggaagcgg cggaggtggg cgcgccgacc cagctttctt gtacaaagtt ggcattataa
     1801 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
     1861 ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt
     1921 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     1981 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     2041 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
     2101 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
     2161 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     2221 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     2281 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa
     2341 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     2401 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     2461 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     2521 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     2581 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     2641 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     2701 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     2761 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     2821 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     2881 gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt
     2941 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3001 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3061 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3121 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3181 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3241 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3301 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3361 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3421 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3481 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3541 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3601 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3661 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3721 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     3781 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     3841 ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg
     3901 tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt
     3961 ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc
     4021 tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
     4081 ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg
     4141 cgttaacgct agcatggatg ttttcccagt cacgacgtt
//
