LOCUS       sequence-246258-        4288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway entry vector for making C-terminal miniTurbo-mVenus fusion 
            with non-nuclear proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4288)
  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 4288)
  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-NES-mVenus-STOP-L3
FEATURES             Location/Qualifiers
     source          1..4288
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             123..164
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             171..941
                     /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             948..974
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             join(1014..1016,1017..1019,1020..1733)
                     /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(1320..1339)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1667..1688
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1734..1829)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1866..1885)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1867..1885)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1890..1906)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1890..1906)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2008..2027)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2019..2828
                     /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(2086..2105)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2975..3563
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3464..3483
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3717..3734
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3893..3920)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4012..4098)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4147..4169
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4161..4178
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4162..4178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(join(4249..4288,1..84))
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc
       61 gttcaacttt cttgtacaaa gtggctgggc gcgccggtgg aggcggttca ggaggtggca
      121 tgggcaagcc catccccaac cccctgctgg gcctggacag caccgctagc atcccgctgc
      181 tgaacgctaa acagattctg ggacagctgg acggcgggag cgtggcagtc ctgcctgtgg
      241 tcgactccac caatcagtac ctgctggatc gaatcggcga gctgaagagt ggggatgctt
      301 gcattgcaga atatcagcag gcagggagag gaagcagagg gaggaaatgg ttctctcctt
      361 ttggagctaa cctgtacctg agtatgtttt ggcgcctgaa gcggggacca gcagcaatcg
      421 gcctgggccc ggtcatcgga attgtcatgg cagaagcgct gcgaaagctg ggagcagaca
      481 aggtgcgagt caaatggccc aatgacctgt atctgcagga tagaaagctg gcaggcatcc
      541 tggtggagct ggccggaata acaggcgatg ctgcacagat cgtcattggc gccgggatta
      601 acgtggctat gaggcgcgtg gaggaaagcg tggtcaatca gggctggatc acactgcagg
      661 aagcagggat taacctggac aggaatactc tggccgctat gctgatccga gagctgcggg
      721 cagccctgga actgttcgag caggaaggcc tggctccata tctgtcacgg tgggagaagc
      781 tggataactt catcaataga cccgtgaagc tgatcattgg ggacaaagag attttcggga
      841 ttagccgggg gattgataaa cagggagccc tgctgctgga acaggacgga gttatcaaac
      901 cctggatggg cggagaaatc agtctgcggt ctgccgaaaa gctgcagctg cctcccctgg
      961 agcgcctgac cctggacgga ggcggtggaa gcggcggagg tgggcgcgcc tccatggtga
     1021 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     1081 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     1141 tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     1201 ccaccctggg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
     1261 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1321 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1381 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1441 agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca
     1501 aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact
     1561 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1621 gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1681 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tgacaacttt
     1741 attatacaaa gttggcatta taaaaaagca ttgcttatca atttgttgca acgaacaggt
     1801 cactatcagt caaaataaaa tcattatttg gagctccatg gtagcgttaa cgcggccgcg
     1861 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     1921 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     1981 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     2041 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     2101 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
     2161 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     2221 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     2281 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
     2341 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     2401 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     2461 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     2521 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     2581 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     2641 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     2701 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     2761 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     2821 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac
     2881 gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag
     2941 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3001 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3061 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3121 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3181 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3241 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3301 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3361 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3421 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3481 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3541 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3601 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3661 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3721 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     3781 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     3841 gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc
     3901 atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc
     3961 ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct
     4021 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg
     4081 agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga
     4141 tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg ccctgcagct
     4201 ctagagctcg aattctacag gtcactaata ccatctaagt agttggttca tagtgactgc
     4261 atatgttgtg ttttacagta ttatgtag
//
