LOCUS       sequence-243657-        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     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(624..747)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             786..827
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             834..1604
                     /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(1641..1643,1644..1646,1647..2360)
                     /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(1947..1966)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2294..2315
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2361..2456)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2493..2512)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2494..2512)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2517..2533)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2517..2533)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2635..2654)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2646..3455
                     /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(2713..2732)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3602..4190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4091..4110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggccctg cagctctaga gctcgaattc tacaggtcac
      601 taataccatc taagtagttg gttcatagtg actgcatatg ttgtgtttta cagtattatg
      661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
      721 ctcgttcaac tttcttgtac aaagtggctg ggcgcgccgg tggaggcggt tcaggaggtg
      781 gcatgggcaa gcccatcccc aaccccctgc tgggcctgga cagcaccgct agcatcccgc
      841 tgctgaacgc taaacagatt ctgggacagc tggacggcgg gagcgtggca gtcctgcctg
      901 tggtcgactc caccaatcag tacctgctgg atcgaatcgg cgagctgaag agtggggatg
      961 cttgcattgc agaatatcag caggcaggga gaggaagcag agggaggaaa tggttctctc
     1021 cttttggagc taacctgtac ctgagtatgt tttggcgcct gaagcgggga ccagcagcaa
     1081 tcggcctggg cccggtcatc ggaattgtca tggcagaagc gctgcgaaag ctgggagcag
     1141 acaaggtgcg agtcaaatgg cccaatgacc tgtatctgca ggatagaaag ctggcaggca
     1201 tcctggtgga gctggccgga ataacaggcg atgctgcaca gatcgtcatt ggcgccggga
     1261 ttaacgtggc tatgaggcgc gtggaggaaa gcgtggtcaa tcagggctgg atcacactgc
     1321 aggaagcagg gattaacctg gacaggaata ctctggccgc tatgctgatc cgagagctgc
     1381 gggcagccct ggaactgttc gagcaggaag gcctggctcc atatctgtca cggtgggaga
     1441 agctggataa cttcatcaat agacccgtga agctgatcat tggggacaaa gagattttcg
     1501 ggattagccg ggggattgat aaacagggag ccctgctgct ggaacaggac ggagttatca
     1561 aaccctggat gggcggagaa atcagtctgc ggtctgccga aaagggaggc ggtggaagcg
     1621 gcggaggtgg gcgcgcctcc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     1681 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1741 gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc
     1801 tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcctgcag tgcttcgccc
     1861 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1921 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1981 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2041 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     2101 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg
     2161 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2221 tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg
     2281 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2341 tggacgagct gtacaagtga caactttatt atacaaagtt ggcattataa aaaagcattg
     2401 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttggag
     2461 ctccatggta gcgttaacgc ggccgcgata tcccctatag tgagtcgtat tacatggtca
     2521 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     2581 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     2641 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg
     2701 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     2761 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     2821 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     2881 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     2941 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     3001 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     3061 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     3121 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     3181 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     3241 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     3301 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     3361 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     3421 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
     3481 actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta
     3541 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3601 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3661 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3721 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     3781 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3841 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3901 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3961 cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag
     4021 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4081 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4141 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4201 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
