LOCUS       sequence-246287-        4355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway entry vector for expressing nuclear BirA* (R118G)-YFP under 
            a promoter of choice.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4355)
  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 4355)
  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)-YFP-NLS-STOP-L2
FEATURES             Location/Qualifiers
     source          1..4355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    16..115
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             129..170
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             363..1133
                     /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             join(1161..1163,1164..1166,1167..1877)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1206..1227)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1467..1486)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1814..1835
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1887..1907
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(1927..2026)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2043..2062)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2044..2062)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2067..2083)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2067..2083)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2185..2204)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2196..3005
                     /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(2263..2282)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3152..3740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3641..3660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3894..3911
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4070..4097)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4189..4275)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4324..4346
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4338..4355
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4339..4355
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cttaagctcg ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa
       61 caaattgatg agcaatgctt ttttataatg ccaactttgt acaaaaaagc aggctccgcg
      121 gccgcatggg caagcccatc cccaaccccc tgctgggcct ggacagcacc gctagcaagg
      181 ataacaccgt gcccctgaaa ctgattgctc tgctggctaa tggcgagttc catagtggcg
      241 aacagctggg agaaaccctg ggcatgtcca gggccgctat caacaagcac attcagactc
      301 tgcgcgactg gggcgtggac gtgttcaccg tgcccggaaa gggctactct ctgcccgagc
      361 ctatccagct gctgaacgct aaacagattc tgggacagct ggacggcggg agcgtggcag
      421 tcctgcctgt gatcgactcc accaatcagt acctgctgga tcgaatcggc gagctgaaga
      481 gtggggatgc ttgcattgca gaatatcagc aggcaggaag aggagggaga gggaggaaat
      541 ggttctctcc ttttggagct aacctgtacc tgagtatgtt ttggcgcctg gagcagggac
      601 cagcagcaat cggcctgagc ctggtcatcg gaattgtcat ggcagaagtg ctgcgaaagc
      661 tgggagcaga caaggtgcga gtcaaatggc ccaatgacct gtatctgcag gatagaaagc
      721 tggcaggcat cctggtggag ctgaccggaa aaacaggcga tgctgcacag atcgtcattg
      781 gcgccgggat taacatggct atgaggcgcg tggaggaaag cgtggtcaat cagggctgga
      841 tcacactgca ggaagcaggg attaacctgg acaggaatac tctggccgct atgctgatcc
      901 gagagctgcg ggcagccctg gaactgttcg agcaggaagg cctggctcca tatctgtcac
      961 ggtgggagaa gctggataac ttcatcaata gacccgtgaa gctgatcatt ggggacaaag
     1021 agattttcgg gattagccgg ggaattgata aacagggagc cctgctgctg gaacaggacg
     1081 gaattatcaa accctggatg ggcggagaaa tcagtctgcg gtctgccgaa aagggaggcg
     1141 gtggaagcgg cggaggttcc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     1201 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1261 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1321 tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc
     1381 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1441 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1501 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1561 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     1621 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     1681 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     1741 tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg
     1801 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1861 tggacgagct gtacaaggct tcggatccaa aaaagaagag aaaggtatag aagggtgggc
     1921 gcgccgaccc agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt
     1981 gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata
     2041 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
     2101 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     2161 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     2221 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     2281 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
     2341 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     2401 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     2461 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
     2521 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     2581 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     2641 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     2701 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     2761 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     2821 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     2881 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     2941 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     3001 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     3061 acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     3121 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3181 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3241 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     3301 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3361 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3421 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3481 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3541 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3601 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3661 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3721 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3781 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3841 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3901 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     3961 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     4021 acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc
     4081 cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg
     4141 ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact
     4201 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc
     4261 ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt
     4321 tttcccagtc acgacgttgt aaaacgacgg ccagt
//
