LOCUS       sequence-170762-        4127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Middle entry vector for generating MBS tagged RNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4127)
  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 4127)
  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-24xMBS
FEATURES             Location/Qualifiers
     source          1..4127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1346..1445)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1461..1478)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1461..1477)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1470..1492)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1541..1627
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1719..1746
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1905..1922)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2076..2664)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2156..2175)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2757..3548)
                     /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"
     primer_bind     3979..3995
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3979..3995
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4000..4019
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4000..4018
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(4036..4127,1..8)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 agctgggtcg aattcgccct tcgcgagatc tgatgaaccc tggaatactg gagcgacgcg
       61 tgatcgtcgt cgtttgaaga ttcgacctgg aggcacggct gatgctcgtg ctttcttggc
      121 aataagtacc gtaggatctg atgaaccctg gaatactgga gcgacgcgtg atcgtcgtcg
      181 tttgaagatt cgacctggag gcacggctga tgctcgtgct ttcttggcaa taagtaccgt
      241 aggatctgat gaaccctgga atactggagc gacgcgtgat cgtcgtcgtt tgaagattcg
      301 acctggaggc acggctgatg ctcgtgcttt cttggcaata agtaccgtag gatctgatga
      361 accctggaat actggagcga cgcgtgatcg tcgtcgtttg aagattcgac ctggaggcac
      421 ggctgatgct cgtgctttct tggcaataag taccgtagga tctgatgaac cctggaatac
      481 tggagcgacg cgtgatcgtc gtcgtttgaa gattcgacct ggaggcacgg ctgatgctcg
      541 tgctttcttg gcaataagta ccgtaggatc tgatgaaccc tggaatactg gagcgacgcg
      601 tgatcgtcgt cgtttgaaga ttcgacctgg aggcacggct gatgctcgtg ctttcttggc
      661 aataagtacc gtaggatctg atgaaccctg gaatactgga gcgacgcgtg atcgtcgtcg
      721 tttgaagatt cgacctggag gcacggctga tgctcgtgct ttcttggcaa taagtaccgt
      781 aggatctgat gaaccctgga atactggagc gacgcgtgat cgtcgtcgtt tgaagattcg
      841 acctggaggc acggctgatg ctcgtgcttt cttggcaata agtaccgtag gatctgatga
      901 accctggaat actggagcga cgcgtgatcg tcgtcgtttg aagattcgac ctggaggcac
      961 ggctgatgct cgtgctttct tggcaataag taccgtagga tctgatgaac cctggaatac
     1021 tggagcgacg cgtgatcgtc gtcgtttgaa gattcgacct ggaggcacgg ctgatgctcg
     1081 tgctttcttg gcaataagta ccgtaggatc tgatgaaccc tggaatactg gagcgacgcg
     1141 tgatcgtcgt cgtttgaaga ttcgacctgg aggcacggct gatgctcgtg ctttcttggc
     1201 aataagtacc gtaggatctg atgaaccctg gaatactgga gcgacgcgtg atcgtcgtcg
     1261 tttgaagatt cgacctggag gcacggctga tgctcgtgct ttcttggcaa taagtaccgt
     1321 aggatcccgc gaagggcgaa ttcggagcct gcttttttgt acaaagttgg cattataaaa
     1381 aagcattgct catcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     1441 atttggggcc cgagcttaag actggccgtc gttttacaac gtcgtgactg ggaaaacatc
     1501 catgctagcg ttaacgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca
     1561 gtcggaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     1621 gacaaatccg ccgggagcgg atttgaacgt tgtgaagcaa cggcccggag ggtggcgggc
     1681 aggacgcccg ccataaactg ccaggcatca aactaagcag aaggccatcc tgacggatgg
     1741 cctttttgcg tttctacaaa ctcttcctgg ctagcggtac gcgtattaat tgcgttgcgc
     1801 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1861 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     1921 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1981 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2041 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2101 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2161 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2221 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2281 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2341 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2401 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2461 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2521 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2581 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2641 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2701 cagtggaacg acgcgtaact cacgttaagg gattttggtc atgggtggct cgagggttat
     2761 ttgccgacta ccttggtgat ctcgcctttc acgtagtgga caaattcttc caactgatct
     2821 gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag cctgtctagc ttcaagtatg
     2881 acgggctgat actgggccgg caggcgctcc attgcccagt cggcagcgac atccttcggc
     2941 gcgattttgc cggttactgc gctgtaccaa atgcgggaca acgtaagcac tacatttcgc
     3001 tcatcgccag cccagtcggg cggcgagttc catagcgtta aggtttcatt tagcgcctca
     3061 aatagatcct gttcaggaac cggatcaaag agttcctccg ccgctggacc taccaaggca
     3121 acgctatgtt ctcttgcttt tgtcagcaag atagccagat caatgtcgat cgtggctggc
     3181 tcgaagatac ctgcaagaat gtcattgcgc tgccattctc caaattgcag ttcgcgctta
     3241 gctggataac gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg
     3301 agaatctcgc tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc
     3361 cgcgttgttt catcaagcct tacggtcacc gtaaccagca aatcaatatc actgtgtggc
     3421 ttcaggccgc catccactgc ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga
     3481 tggcgctcga tgacgccaac tacctctgat agttgagtcg atacttcggc gatcaccgct
     3541 tccctcataa tgtttaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg
     3601 ctccataaca tcaaacatcg acccacggcg taacgcgctt gctgcttgga tgcccgaggc
     3661 atagactgta ccccaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta
     3721 ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat
     3781 tacagcttac gaaccgaaca ggcttatgtc cactgggttc gtgccttcat ccgtttccac
     3841 ggtgtgcgtc acccggcaac cttgggtagc agcgaagtcg aggcatttct gtcctggctg
     3901 gtctagagga ggcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga
     3961 gacacgggcc agagctgcca ggaaacagct atgaccatgt aatacgactc actatagggg
     4021 atatcagctg gatggcaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa
     4081 caaattgatg agcaattatt ttttataatg ccaactttgt acaagaa
//
