LOCUS       sequence-246285-        4252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway entry vector for making C-terminal miniTurbo-mVenus fusion 
            with nuclear and non-nuclear proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4252)
  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 4252)
  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: pDONR_P2R-P3_R2-miniTurbo-mVenus-STOP-L3
FEATURES             Location/Qualifiers
     source          1..4252
                     /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    complement(88..211)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             250..291
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             298..1068
                     /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(1105..1107,1108..1110,1111..1824)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1411..1430)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1758..1779
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1825..1920)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1957..1976)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1958..1976)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1981..1997)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1981..1997)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2099..2118)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2110..2919
                     /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(2177..2196)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3066..3654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3555..3574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3808..3825
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3984..4011)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4103..4189)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4238..4252,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4252,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cctgcagctc tagagctcga attctacagg
       61 tcactaatac catctaagta gttggttcat agtgactgca tatgttgtgt tttacagtat
      121 tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat atcattttac
      181 gtttctcgtt caactttctt gtacaaagtg gctgggcgcg ccggtggagg cggttcagga
      241 ggtggcatgg gcaagcccat ccccaacccc ctgctgggcc tggacagcac cgctagcatc
      301 ccgctgctga acgctaaaca gattctggga cagctggacg gcgggagcgt ggcagtcctg
      361 cctgtggtcg actccaccaa tcagtacctg ctggatcgaa tcggcgagct gaagagtggg
      421 gatgcttgca ttgcagaata tcagcaggca gggagaggaa gcagagggag gaaatggttc
      481 tctccttttg gagctaacct gtacctgagt atgttttggc gcctgaagcg gggaccagca
      541 gcaatcggcc tgggcccggt catcggaatt gtcatggcag aagcgctgcg aaagctggga
      601 gcagacaagg tgcgagtcaa atggcccaat gacctgtatc tgcaggatag aaagctggca
      661 ggcatcctgg tggagctggc cggaataaca ggcgatgctg cacagatcgt cattggcgcc
      721 gggattaacg tggctatgag gcgcgtggag gaaagcgtgg tcaatcaggg ctggatcaca
      781 ctgcaggaag cagggattaa cctggacagg aatactctgg ccgctatgct gatccgagag
      841 ctgcgggcag ccctggaact gttcgagcag gaaggcctgg ctccatatct gtcacggtgg
      901 gagaagctgg ataacttcat caatagaccc gtgaagctga tcattgggga caaagagatt
      961 ttcgggatta gccgggggat tgataaacag ggagccctgc tgctggaaca ggacggagtt
     1021 atcaaaccct ggatgggcgg agaaatcagt ctgcggtctg ccgaaaaggg aggcggtgga
     1081 agcggcggag gtgggcgcgc ctccatggtg agcaagggcg aggagctgtt caccggggtg
     1141 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1201 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc
     1261 aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcct gcagtgcttc
     1321 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1381 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1441 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1501 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1561 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     1621 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1681 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc
     1741 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1801 ggcatggacg agctgtacaa gtgacaactt tattatacaa agttggcatt ataaaaaagc
     1861 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     1921 ggagctccat ggtagcgtta acgcggccgc gatatcccct atagtgagtc gtattacatg
     1981 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     2041 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     2101 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     2161 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     2221 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     2281 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     2341 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     2401 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     2461 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     2521 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     2581 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     2641 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     2701 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     2761 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     2821 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2881 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     2941 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     3001 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3061 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3121 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3181 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3241 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3301 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3361 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3421 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3481 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3541 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3601 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3661 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3721 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3781 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3841 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     3901 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc
     3961 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt
     4021 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa
     4081 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     4141 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
     4201 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg tt
//
