LOCUS       sequence-243663-        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    complement(17..116)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(133..152)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(134..152)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(157..173)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(157..173)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(275..294)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             286..1095
                     /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(353..372)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1242..1830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1731..1750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1984..2001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2160..2187)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2279..2365)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2414..2436
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2428..2445
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2429..2445
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2461..2560
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2574..2615
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2622..3392
                     /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(3420..3422,3423..3425,3426..4136)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3726..3745)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4073..4094
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4146..4166
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 aagggtgggc gcgccgaccc agctttcttg tacaaagttg gcattataag aaagcattgc
       61 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
      121 ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
      181 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
      241 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
      301 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
      361 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
      421 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
      481 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
      541 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
      601 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
      661 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
      721 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
      781 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
      841 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
      901 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
      961 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     1021 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     1081 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     1141 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc
     1201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1381 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1621 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     1801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     1861 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     1921 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     1981 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     2041 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     2101 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa
     2161 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
     2221 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
     2281 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     2341 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
     2401 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
     2461 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
     2521 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgcggccgc atgggcaagc
     2581 ccatccccaa ccccctgctg ggcctggaca gcaccgctag catcccgctg ctgaacgcta
     2641 aacagattct gggacagctg gacggcggga gcgtggcagt cctgcctgtg gtcgactcca
     2701 ccaatcagta cctgctggat cgaatcggcg agctgaagag tggggatgct tgcattgcag
     2761 aatatcagca ggcagggaga ggaagcagag ggaggaaatg gttctctcct tttggagcta
     2821 acctgtacct gagtatgttt tggcgcctga agcggggacc agcagcaatc ggcctgggcc
     2881 cggtcatcgg aattgtcatg gcagaagcgc tgcgaaagct gggagcagac aaggtgcgag
     2941 tcaaatggcc caatgacctg tatctgcagg atagaaagct ggcaggcatc ctggtggagc
     3001 tggccggaat aacaggcgat gctgcacaga tcgtcattgg cgccgggatt aacgtggcta
     3061 tgaggcgcgt ggaggaaagc gtggtcaatc agggctggat cacactgcag gaagcaggga
     3121 ttaacctgga caggaatact ctggccgcta tgctgatccg agagctgcgg gcagccctgg
     3181 aactgttcga gcaggaaggc ctggctccat atctgtcacg gtgggagaag ctggataact
     3241 tcatcaatag acccgtgaag ctgatcattg gggacaaaga gattttcggg attagccggg
     3301 ggattgataa acagggagcc ctgctgctgg aacaggacgg agttatcaaa ccctggatgg
     3361 gcggagaaat cagtctgcgg tctgccgaaa agggaggcgg tggaagcggc ggaggttcca
     3421 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     3481 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     3541 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     3601 tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc
     3661 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3721 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3781 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3841 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     3901 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     3961 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     4021 acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     4081 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaaggctt
     4141 cggatccaaa aaagaagaga aaggtatag
//
