LOCUS       sequence-243650-        4397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway entry vector for making N-terminal YFP-TurboID fusions with 
            non-nuclear proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4397)
  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 4397)
  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-Turbo-NES-L2
FEATURES             Location/Qualifiers
     source          1..4397
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             763..804
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             811..1767
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             1774..1800
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     protein_bind    complement(1838..1937)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1954..1973)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1955..1973)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1978..1994)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1978..1994)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2096..2115)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2107..2916
                     /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(2174..2193)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3063..3651
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3552..3571
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3805..3822
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3981..4008)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4100..4186)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4235..4257
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4249..4266
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4250..4266
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4282..4381
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      481 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaaggct
      721 aagggtgggc gcgccggtgg aggcggttca ggaggtggca tgggcaagcc catccccaac
      781 cccctgctgg gcctggacag caccgctagc aaagacaata ctgtgcctct gaagctgatc
      841 gctctcctgg ctaatggcga gttccatagt ggcgaacagc tgggagaaac cctgggcatg
      901 tccagggccg ctatcaacaa gcacattcag actctgcgcg actggggcgt ggacgtgttc
      961 accgtgcccg gaaagggcta ctctctgccc gagcctatcc cgctgctgaa cgctaaacag
     1021 attctgggac agctggacgg cgggagcgtg gcagtcctgc ctgtggtcga ctccaccaat
     1081 cagtacctgc tggatcgaat cggcgagctg aagagtgggg atgcttgcat tgcagaatat
     1141 cagcaggcag ggagaggaag cagagggagg aaatggttct ctccttttgg agctaacctg
     1201 tacctgagta tgttttggcg cctgaagcgg ggaccagcag caatcggcct gggcccggtc
     1261 atcggaattg tcatggcaga agcgctgcga aagctgggag cagacaaggt gcgagtcaaa
     1321 tggcccaatg acctgtatct gcaggataga aagctggcag gcatcctggt ggagctggcc
     1381 ggaataacag gcgatgctgc acagatcgtc attggcgccg ggattaacgt ggctatgagg
     1441 cgcgtggagg aaagcgtggt caatcagggc tggatcacac tgcaggaagc agggattaac
     1501 ctggacagga atactctggc cgctacgctg atccgagagc tgcgggcagc cctggaactg
     1561 ttcgagcagg aaggcctggc tccatatctg ccacggtggg agaagctgga taacttcatc
     1621 aatagacccg tgaagctgat cattggggac aaagagattt tcgggattag ccgggggatt
     1681 gataaacagg gagccctgct gctggaacag gacggagtta tcaaaccctg gatgggcgga
     1741 gaaatcagtc tgcggtctgc cgaaaagctg cagctgcctc ccctggagcg cctgaccctg
     1801 gacggaggcg gtggaagcgg cggaggtggg cgcgccgacc cagctttctt gtacaaagtt
     1861 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     1921 aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc
     1981 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     2041 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     2101 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     2161 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     2221 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     2281 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     2341 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     2401 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     2461 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     2521 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     2581 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     2641 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     2701 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     2761 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     2821 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     2881 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     2941 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
     3001 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3061 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3121 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3181 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     3241 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3301 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3361 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3421 acctacaccg aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa
     3481 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3541 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3601 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3661 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3721 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3781 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     3841 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     3901 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag
     3961 gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat
     4021 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt
     4081 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
     4141 taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc
     4201 cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg
     4261 gccagtctta agctcgggcc ccaaataatg attttatttt gactgatagt gacctgttcg
     4321 ttgcaacaaa ttgatgagca atgctttttt ataatgccaa ctttgtacaa aaaagcaggc
     4381 tccgcggccg ccccctt
//
