LOCUS       sequence-342338-        3355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite Gateway middle entry clone encoding miniTurbo. Parton lab 
            clone JRQ.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3355)
  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 3355)
  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-miniTurbo
FEATURES             Location/Qualifiers
     source          1..3355
                     /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(1480..1579)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1596..1615)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1597..1615)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1620..1636)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1620..1636)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1738..1757)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1749..2558
                     /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(1816..1835)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2705..3293
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3194..3213
                     /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 caccatggaa caaaagctca tctcagaaga agatcttatt ccattgttga
      721 acgcgaagca aattctcggc caacttgatg gggggagcgt tgcggtgctc cccgttgtgg
      781 atagcactaa tcaatacctt ctcgatagga ttggcgagct gaaatccggc gacgcctgta
      841 ttgcagagta ccagcaggcc gggcgggggt ctcgagggcg taagtggttc tcaccgttcg
      901 gtgcgaacct ctacctctca atgttttggc gattgaagag ggggccggca gctatcgggt
      961 tggggccagt gatagggatt gttatggcag aggcgctcag gaaactcggc gcggacaagg
     1021 tccgcgtgaa atggccaaac gatttgtacc ttcaagaccg gaagttggca ggaattctgg
     1081 tggagctcgc cggaataacc ggagatgccg cacaaatagt cattggtgca gggataaatg
     1141 ttgcgatgcg tcgggtcgag gaaagcgttg taaaccaagg gtggattaca ttgcaggagg
     1201 cgggcatcaa tttggataga aatactcttg ctgcaatgtt gataagggag ttgcgcgcgg
     1261 ccttggaatt gttcgagcag gaaggcttgg ccccgtattt gagtaggtgg gagaaactgg
     1321 acaattttat taaccgccca gttaagttga tcatcggcga taaagaaatt ttcggaatat
     1381 ccagaggtat tgacaagcaa ggcgcgctcc tcctggagca ggatggggtg atcaagccgt
     1441 ggatgggtgg tgaaatctcc ctgagaagcg cggaaaaaca cccagctttc ttgtacaaag
     1501 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     1561 aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg
     1621 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     1681 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     1741 ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg
     1801 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     1861 tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     1921 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     1981 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     2041 cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     2101 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     2161 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     2221 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     2281 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     2341 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     2401 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     2461 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     2521 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt
     2581 aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc
     2641 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2701 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2761 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2821 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     2881 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2941 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3001 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3061 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3121 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3181 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3241 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3301 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
