LOCUS       sequence-342325-        3541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite Gateway middle entry clone encoding TurboID. Parton lab 
            clone JRP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3541)
  AUTHORS   Xiong Z, Lo HP, McMahon KA, Martel N, Jones A, Hill MM, Parton RG, 
            Hall TE
  TITLE     In vivo proteomic mapping through GFP-directed proximity-dependent 
            biotin labelling in zebrafish.
  JOURNAL   Elife. 2021 Feb 16;10. pii: 64631. doi: 10.7554/eLife.64631.
  PUBMED    33591275
REFERENCE   2  (bases 1 to 3541)
  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: "2021-02-16"; volume: "10. pii: 64631. doi"; pages: " 
            10.7554/eLife.64631"
COMMENT     Sequence Label: pME-TurboID-NS
FEATURES             Location/Qualifiers
     source          1..3541
                     /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     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     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"
     regulatory      669..678
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             678..707
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(1666..1765)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1782..1801)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1783..1801)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1806..1822)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1806..1822)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1924..1943)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1935..2744
                     /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(2002..2021)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2891..3479
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3380..3399
                     /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 agcaggctgc caccatggag cagaaactga taagcgaaga ggacctgaaa gacaatacag
      721 tccccttgaa gctgattgcc cttctggcaa atggcgaatt ccacagcggg gagcaactgg
      781 gcgaaacgtt gggcatgagt agagcagcga taaacaaaca tatacaaacg ctgagggatt
      841 ggggtgtaga cgtgttcacc gttccgggca agggctatag tttgcccgaa ccgataccac
      901 ttctgaacgc caaacaaata ttgggtcaac tggatggagg atcagtagca gttcttcctg
      961 tggtcgactc tactaaccaa tatctgctgg atcggatcgg ggaactgaaa tctggcgatg
     1021 catgcatagc tgagtaccag caggctggac ggggatctcg aggaaggaaa tggttttccc
     1081 cgtttggggc caacctctat ctctctatgt tctggaggtt gaaacgtggc cctgctgcca
     1141 tcggcttggg gcctgtcatt ggtattgtta tggcggaggc gttgcgtaaa cttggagcgg
     1201 ataaagtgag ggttaaatgg ccaaacgatt tgtatcttca agaccgcaag ttggctggta
     1261 tactggtaga actcgcaggg attacaggag atgcggctca gattgtgatc ggtgccggta
     1321 ttaacgtcgc gatgcgtagg gtagaggagt ccgtggtcaa tcagggttgg attacgctgc
     1381 aagaggctgg aatcaatctc gacagaaaca cacttgcagc cacgcttatt cgggaattga
     1441 gggctgctct tgaacttttt gaacaggaag ggttggcacc ttacctgccg cgctgggaaa
     1501 agctggataa tttcataaat cggcccgtca agctcataat cggggacaag gaaattttcg
     1561 gcatatcacg cggcatcgac aagcaggggg cactgctgtt ggaacaggac ggagtcataa
     1621 agccctggat gggtggtgag attagccttc gatctgcgga gaaacaccca gctttcttgt
     1681 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta
     1741 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     1801 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     1861 ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta
     1921 atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
     1981 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     2041 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     2101 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     2161 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     2221 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     2281 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     2341 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     2401 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     2461 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     2521 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     2581 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     2641 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     2701 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     2761 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
     2821 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     2881 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2941 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3001 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3061 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3121 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3181 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3241 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3301 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3361 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3421 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3481 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3541 t
//
