LOCUS       sequence-340315-        3645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Promiscuous biotin ligase (BirA-G3) targeted to ER lumen in Gateway 
            entry vector. N-terminal secretion signal from Drosophila BiP. 
            C-terminal KDEL. Myc tag inserted after sig. seq. cleavage site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3645)
  AUTHORS   Droujinine IA, Meyer AS, Wang D, Udeshi ND, Hu Y, Rocco D, McMahon 
            JA, Yang R, Guo J, Mu L, Carey DK, Svinkina T, Zeng R, Branon T, 
            Tabatabai A, Bosch JA, Asara JM, Ting AY, Carr SA, McMahon AP, 
            Perrimon N
  TITLE     Proteomics of protein trafficking by in vivo tissue-specific 
            labeling.
  JOURNAL   Nat Commun. 2021 Apr 22;12(1):2382. doi: 10.1038/s41467-021-22599-x.
  PUBMED    33888706
REFERENCE   2  (bases 1 to 3645)
  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: "Nat 
            Commun."; date: "2021-04-22"; volume: "12(1):2382. doi"; pages: " 
            10.1038/s41467-021-22599-x"
COMMENT     Sequence Label: pEntr_Myc-BirA-G3-ER
FEATURES             Location/Qualifiers
     source          1..3645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     sig_peptide     689..742
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             743..772
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             779..1735
                     /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
                     ADKVRVKWPNDLYLQDRKLAGILVELAGKTGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     protein_bind    complement(1770..1869)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1886..1905)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1887..1905)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1910..1926)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1910..1926)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2028..2047)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2039..2848
                     /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(2106..2125)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2995..3583
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3484..3503
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gcggccgccc ccttcaccat gaagttatgc atattactgg ccgtcgtggc
      721 ctttgttggc ctctcgctcg gggaacaaaa gcttatttct gaagaggact tggctagcaa
      781 agacaatact gtgcctctga agctgatcgc tctcctggct aatggcgagt tccatagtgg
      841 cgaacagctg ggagaaaccc tgggcatgtc cagggccgct atcaacaagc acattcagac
      901 tctgcgcgac tggggcgtgg acgtgttcac cgtgcccgga aagggctact ctctgcccga
      961 gcctatcccg ctgctgaacg ctaaacagat tctgggacag ctggacggcg ggagcgtggc
     1021 agtcctgcct gtggtcgact ccaccaatca gtacctgctg gatcgaatcg gcgagctgaa
     1081 gagtggggat gcttgcattg cagaatatca gcaggcaggg agaggaagca gagggaggaa
     1141 atggttctct ccttttggag ctaacctgta cctgagtatg ttttggcgcc tgaagcgggg
     1201 accagcagca atcggcctgg gcccggtcat cggaattgtc atggcagaag cgctgcgaaa
     1261 gctgggagca gacaaggtgc gagtcaaatg gcccaatgac ctgtatctgc aggatagaaa
     1321 gctggcaggc atcctggtgg agctggccgg aaaaacaggc gatgctgcac agatcgtcat
     1381 tggcgccggg attaacgtgg ctatgaggcg cgtggaggaa agcgtggtca atcagggctg
     1441 gatcacactg caggaagcag ggattaacct ggacaggaat actctggccg ctatgctgat
     1501 ccgagagctg cgggcagccc tggaactgtt cgagcaggaa ggcctggctc catatctgtc
     1561 acggtgggag aagctggata acttcatcaa tagacccgtg aagctgatca ttggggacaa
     1621 agagattttc gggattagcc gggggattga taaacaggga gccctgctgc tggaacagga
     1681 cggagttatc aaaccctgga tgggcggaga aatcagtctg cggtctgccg aaaagaagga
     1741 cgagctgtaa tagaagggtg ggcgcgccga cccagctttc ttgtacaaag ttggcattat
     1801 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     1861 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     1921 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     1981 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     2041 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     2101 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     2161 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     2221 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     2281 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     2341 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     2401 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     2461 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     2521 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     2581 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     2641 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     2701 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     2761 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     2821 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     2881 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     2941 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3001 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3061 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3121 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     3181 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3241 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3301 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3361 cgaactgaga tacctacagc gtgagcattg agaaagcgcc acgcttcccg aagggagaaa
     3421 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3481 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3541 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3601 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
