LOCUS       sequence-170769-        3872 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Middle entry vector for generating MBS tagged RNAs with a 
            fluorescent protein reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3872)
  AUTHORS   Campbell PD, Chao JA, Singer RH, Marlow FL
  TITLE     Dynamic visualization of transcription and RNA subcellular 
            localization in zebrafish.
  JOURNAL   Development. 2015 Apr 1;142(7):1368-74. doi: 10.1242/dev.118968. 
            Epub 2015 Mar 10.
  PUBMED    25758462
REFERENCE   2  (bases 1 to 3872)
  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"; doi: 
            "10.1242/dev.118968"; journalName: "Development"; date: "2015-04-1- 
            1"; volume: "142"; issue: "7"; pages: "1368-74"
COMMENT     Sequence Label: pME-mCherry-24xMBS
FEATURES             Location/Qualifiers
     source          1..3872
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    complement(348..447)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(464..483)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(465..483)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(488..504)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(488..504)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             935..1726
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1819..2407
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2308..2327
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2561..2578
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2737..2764)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2856..2942)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2991..3013
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3005..3022
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3006..3022
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3038..3137
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3162..3872
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEDDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3307..3325)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3575..3594
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3769..3792
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
ORIGIN
        1 ggatcctacg gtacttattg ccaagaaagc acgagcatca gccgtgcctc caggtcgaat
       61 cttcaaacga cgacgatcac gcgtcgctcc agtattccag ggttcatcag atcctacggt
      121 acttattgcc aagaaagcac gagcatcagc cgtgcctcca ggtcgaatct tcaaacgacg
      181 acgatcacgc gtcgctccag tattccaggg ttcatcagat cctacggtac ttattgccaa
      241 gaaagcacga gcatcagccg tgcctccagg tcgaatcttc aaacgacgac gatcacgcgt
      301 cgctccagta ttccagggtt catcagatct cgcgaagggc gaattcgacc cagctttctt
      361 gtacaaagtt ggcattataa aaaataattg ctcatcaatt tgttgcaacg aacaggtcac
      421 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
      481 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
      541 cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg acagaaatgc
      601 ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc
      661 acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg
      721 ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt
      781 tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca
      841 gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca
      901 gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt
      961 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     1021 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
     1081 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     1141 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     1201 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     1261 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
     1321 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     1381 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
     1441 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     1501 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     1561 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     1621 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     1681 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct cgagccaccc
     1741 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     1801 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1861 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1921 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1981 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2041 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2101 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2161 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2221 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2281 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2341 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2401 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2461 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2521 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2581 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     2641 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac
     2701 gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
     2761 ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
     2821 cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
     2881 accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt
     2941 tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg
     3001 acgttgtaaa acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact
     3061 gatagtgacc tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt
     3121 gtacaaaaaa gcaggctccg aattcgccct tcgcgggatc catggtgagc aagggcgagg
     3181 acgacaacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg
     3241 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     3301 agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt
     3361 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
     3421 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     3481 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
     3541 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
     3601 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
     3661 agcagaggct gaagctgaag gacggcggcc actacgacgc cgaggtcaag accacctaca
     3721 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ctggacatca
     3781 cctcccacaa cgaggactac accatcgtgg aacagtacga gcgcgccgag ggccgccact
     3841 ccaccggcgg catggacgag ctgtacaagt aa
//
