LOCUS       sequence-246289-        4215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway entry vector for making N-terminal YFP-miniTurbo fusions 
            with non-nuclear proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4215)
  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 4215)
  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-YFP-miniTurbo-NES-L2
FEATURES             Location/Qualifiers
     source          1..4215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(204..223)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     551..572
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             660..701
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             708..1478
                     /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             1485..1511
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     protein_bind    complement(1549..1648)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1665..1684)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1666..1684)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1689..1705)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1689..1705)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1807..1826)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1818..2627
                     /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(1885..1904)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2774..3362
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3263..3282
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3516..3533
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3692..3719)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3811..3897)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3946..3968
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3960..3977
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3961..3977
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3993..4092
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(4113..4115,4116..4118,4119..4215,1..614)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
ORIGIN
        1 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
       61 ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct
      121 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
      181 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
      241 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
      301 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
      361 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
      421 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
      481 tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga
      541 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
      601 acgagctgta caaggctaag ggtgggcgcg ccggtggagg cggttcagga ggtggcatgg
      661 gcaagcccat ccccaacccc ctgctgggcc tggacagcac cgctagcatc ccgctgctga
      721 acgctaaaca gattctggga cagctggacg gcgggagcgt ggcagtcctg cctgtggtcg
      781 actccaccaa tcagtacctg ctggatcgaa tcggcgagct gaagagtggg gatgcttgca
      841 ttgcagaata tcagcaggca gggagaggaa gcagagggag gaaatggttc tctccttttg
      901 gagctaacct gtacctgagt atgttttggc gcctgaagcg gggaccagca gcaatcggcc
      961 tgggcccggt catcggaatt gtcatggcag aagcgctgcg aaagctggga gcagacaagg
     1021 tgcgagtcaa atggcccaat gacctgtatc tgcaggatag aaagctggca ggcatcctgg
     1081 tggagctggc cggaataaca ggcgatgctg cacagatcgt cattggcgcc gggattaacg
     1141 tggctatgag gcgcgtggag gaaagcgtgg tcaatcaggg ctggatcaca ctgcaggaag
     1201 cagggattaa cctggacagg aatactctgg ccgctatgct gatccgagag ctgcgggcag
     1261 ccctggaact gttcgagcag gaaggcctgg ctccatatct gtcacggtgg gagaagctgg
     1321 ataacttcat caatagaccc gtgaagctga tcattgggga caaagagatt ttcgggatta
     1381 gccgggggat tgataaacag ggagccctgc tgctggaaca ggacggagtt atcaaaccct
     1441 ggatgggcgg agaaatcagt ctgcggtctg ccgaaaagct gcagctgcct cccctggagc
     1501 gcctgaccct ggacggaggc ggtggaagcg gcggaggtgg gcgcgccgac ccagctttct
     1561 tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca
     1621 ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt
     1681 attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt
     1741 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca
     1801 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt
     1861 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     1921 gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     1981 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     2041 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     2101 tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag
     2161 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     2221 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     2281 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     2341 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg
     2401 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     2461 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     2521 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     2581 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta
     2641 attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac
     2701 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga
     2761 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2821 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2881 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2941 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3001 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3061 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3121 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3181 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3241 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3301 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3361 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3421 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3481 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3541 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     3601 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta
     3661 ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg
     3721 cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg
     3781 cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga
     3841 caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg
     3901 cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt
     3961 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
     4021 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
     4081 aaaaagcagg ctccgcggcc gcccccttca ccatggtgag caagggcgag gagctgttca
     4141 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     4201 tgtccggcga gggcg
//
