LOCUS       sequence-342348-        3262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite Gateway middle entry clone encoding mRuby2. Parton lab 
            clone DJA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3262)
  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 3262)
  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-Ruby2-NS
FEATURES             Location/Qualifiers
     source          1..3262
                     /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             675..1385
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     protein_bind    complement(1387..1486)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1503..1522)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1504..1522)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1527..1543)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1527..1543)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1645..1664)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1656..2465
                     /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(1723..1742)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2612..3200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3101..3120
                     /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 caccatggtg tctaagggcg aagagctgat caaggaaaat atgcgtatga
      721 aggtggtcat ggaaggttcg gtcaacggcc accaattcaa atgcacaggt gaaggagaag
      781 gcaatccgta catgggaact caaaccatga ggatcaaagt catcgaggga ggacccctgc
      841 catttgcctt tgacattctt gccacgtcgt tcatgtatgg cagccgtact tttatcaagt
      901 acccgaaagg cattcctgat ttctttaaac agtcctttcc tgagggtttt acttgggaaa
      961 gagttacgag atacgaagat ggtggagtcg tcaccgtcat gcaggacacc agccttgagg
     1021 atggctgtct cgtttaccac gtccaagtca gaggggtaaa ctttccctcc aatggtcccg
     1081 tgatgcagaa gaagaccaag ggttgggagc ctaatacaga gatgatgtat ccagcagatg
     1141 gtggtctgag gggatacact catatggcac tgaaagttga tggtggtggc catctgtctt
     1201 gctctttcgt aacaacttac aggtcaaaaa agaccgtcgg gaacatcaag atgcccggta
     1261 tccatgccgt tgatcaccgc ctggaaaggt tagaggaaag tgacaatgaa atgttcgtag
     1321 tacaacgcga acacgcagtt gccaagttcg ccgggcttgg tggtgggatg gacgagctgt
     1381 acaagcaccc agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt
     1441 gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata
     1501 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
     1561 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     1621 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     1681 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     1741 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
     1801 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     1861 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     1921 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
     1981 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     2041 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     2101 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     2161 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     2221 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     2281 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     2341 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     2401 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     2461 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     2521 acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     2581 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2641 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2701 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2761 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2821 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2881 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2941 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3001 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3061 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3121 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3181 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3241 gctggccttt tgctcacatg tt
//
