LOCUS       sequence-246307-        4169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway entry vector for expressing nuclear miniTurbo-YFP under a 
            promoter of choice.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4169)
  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 4169)
  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-miniTurbo-YFP-NLS-STOP-L2
FEATURES             Location/Qualifiers
     source          1..4169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    16..115
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             129..170
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             177..947
                     /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"
     CDS             join(975..977,978..980,981..1691)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1281..1300)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1628..1649
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1701..1721
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(1741..1840)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1857..1876)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1858..1876)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1881..1897)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1881..1897)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1999..2018)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2010..2819
                     /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(2077..2096)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2966..3554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3455..3474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3708..3725
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3884..3911)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4003..4089)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4138..4160
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4152..4169
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4153..4169
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cttaagctcg ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa
       61 caaattgatg agcaatgctt ttttataatg ccaactttgt acaaaaaagc aggctccgcg
      121 gccgcatggg caagcccatc cccaaccccc tgctgggcct ggacagcacc gctagcatcc
      181 cgctgctgaa cgctaaacag attctgggac agctggacgg cgggagcgtg gcagtcctgc
      241 ctgtggtcga ctccaccaat cagtacctgc tggatcgaat cggcgagctg aagagtgggg
      301 atgcttgcat tgcagaatat cagcaggcag ggagaggaag cagagggagg aaatggttct
      361 ctccttttgg agctaacctg tacctgagta tgttttggcg cctgaagcgg ggaccagcag
      421 caatcggcct gggcccggtc atcggaattg tcatggcaga agcgctgcga aagctgggag
      481 cagacaaggt gcgagtcaaa tggcccaatg acctgtatct gcaggataga aagctggcag
      541 gcatcctggt ggagctggcc ggaataacag gcgatgctgc acagatcgtc attggcgccg
      601 ggattaacgt ggctatgagg cgcgtggagg aaagcgtggt caatcagggc tggatcacac
      661 tgcaggaagc agggattaac ctggacagga atactctggc cgctatgctg atccgagagc
      721 tgcgggcagc cctggaactg ttcgagcagg aaggcctggc tccatatctg tcacggtggg
      781 agaagctgga taacttcatc aatagacccg tgaagctgat cattggggac aaagagattt
      841 tcgggattag ccgggggatt gataaacagg gagccctgct gctggaacag gacggagtta
      901 tcaaaccctg gatgggcgga gaaatcagtc tgcggtctgc cgaaaaggga ggcggtggaa
      961 gcggcggagg ttccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1021 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1081 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1141 tgccctggcc caccctcgtg accaccttcg gctacggcct gcagtgcttc gcccgctacc
     1201 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1261 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1321 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1381 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     1441 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     1501 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1561 tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc
     1621 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1681 agctgtacaa ggcttcggat ccaaaaaaga agagaaaggt atagaagggt gggcgcgccg
     1741 acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca
     1801 acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct
     1861 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa
     1921 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
     1981 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc
     2041 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     2101 tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt
     2161 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     2221 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     2281 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat
     2341 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     2401 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     2461 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     2521 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     2581 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     2641 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     2701 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     2761 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     2821 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg
     2881 caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     2941 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3001 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3061 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3121 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3181 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3241 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3301 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3361 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3421 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3481 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3541 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3601 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3661 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3721 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     3781 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     3841 ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag
     3901 gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc
     3961 tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag
     4021 agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg
     4081 ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc
     4141 agtcacgacg ttgtaaaacg acggccagt
//
