LOCUS       sequence-246286-        4367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway entry vector for expressing cytosolic BirA* (R118G)-YFP 
            under a promoter of choice.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4367)
  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 4367)
  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-BirA(R118G)-NES-YFP-STOP-L2
FEATURES             Location/Qualifiers
     source          1..4367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..42
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             235..1005
                     /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="IQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGDACI
                     AEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAIGLSLVIGIVMAEVLRKLGADK
                     VRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGWITLQ
                     EAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIF
                     GISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             1012..1038
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             join(1069..1071,1072..1074,1075..1788)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1114..1135)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1375..1394)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1722..1743
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1811..1910)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1927..1946)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1928..1946)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1951..1967)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1951..1967)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2069..2088)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2080..2889
                     /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(2147..2166)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3036..3624
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3525..3544
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3778..3795
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3954..3981)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4073..4159)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4208..4230
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4222..4239
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4223..4239
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4255..4354
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ggcaagccca tccccaaccc cctgctgggc ctggacagca ccgctagcaa ggataacacc
       61 gtgcccctga aactgattgc tctgctggct aatggcgagt tccatagtgg cgaacagctg
      121 ggagaaaccc tgggcatgtc cagggccgct atcaacaagc acattcagac tctgcgcgac
      181 tggggcgtgg acgtgttcac cgtgcccgga aagggctact ctctgcccga gcctatccag
      241 ctgctgaacg ctaaacagat tctgggacag ctggacggcg ggagcgtggc agtcctgcct
      301 gtgatcgact ccaccaatca gtacctgctg gatcgaatcg gcgagctgaa gagtggggat
      361 gcttgcattg cagaatatca gcaggcagga agaggaggga gagggaggaa atggttctct
      421 ccttttggag ctaacctgta cctgagtatg ttttggcgcc tggagcaggg accagcagca
      481 atcggcctga gcctggtcat cggaattgtc atggcagaag tgctgcgaaa gctgggagca
      541 gacaaggtgc gagtcaaatg gcccaatgac ctgtatctgc aggatagaaa gctggcaggc
      601 atcctggtgg agctgaccgg aaaaacaggc gatgctgcac agatcgtcat tggcgccggg
      661 attaacatgg ctatgaggcg cgtggaggaa agcgtggtca atcagggctg gatcacactg
      721 caggaagcag ggattaacct ggacaggaat actctggccg ctatgctgat ccgagagctg
      781 cgggcagccc tggaactgtt cgagcaggaa ggcctggctc catatctgtc acggtgggag
      841 aagctggata acttcatcaa tagacccgtg aagctgatca ttggggacaa agagattttc
      901 gggattagcc ggggaattga taaacaggga gccctgctgc tggaacagga cggaattatc
      961 aaaccctgga tgggcggaga aatcagtctg cggtctgccg aaaagctgca gctgcctccc
     1021 ctggagcgcc tgaccctgga cggaggcggt ggaagcggcg gaggttccat ggtgagcaag
     1081 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1141 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1201 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1261 ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     1321 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1381 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1441 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1501 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1561 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1621 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
     1681 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1741 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagg gcccaagggt
     1801 gggcgcgccg acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
     1861 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
     1921 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     1981 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     2041 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     2101 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     2161 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     2221 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     2281 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     2341 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     2401 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     2461 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     2521 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     2581 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     2641 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     2701 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     2761 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     2821 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     2881 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     2941 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     3001 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3061 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3121 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3181 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3241 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3301 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3361 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3421 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3481 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3541 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3601 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3661 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3721 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3781 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     3841 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     3901 cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc
     3961 catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg
     4021 cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc
     4081 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact
     4141 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg
     4201 atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg gccccaaata
     4261 atgattttat tttgactgat agtgacctgt tcgttgcaac aaattgatga gcaatgcttt
     4321 tttataatgc caactttgta caaaaaagca ggctccgcgg ccgcatg
//
