LOCUS       sequence-406384-        3971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DEST vector for MultiSite Gateway assembly and production of 
            recombinant baculovirus using MultiMate assembly. After LR propagate
            in Pir+. Works as a CRE-donor in the MultiBac system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3971)
  AUTHORS   Aulicino F, Pelosse M, Toelzer C, Capin J, Ilegems E, Meysami P, 
            Rollarson R, Berggren PO, Dillingham MS, Schaffitzel C, Saleem MA, 
            Welsh GI, Berger I
  TITLE     Highly efficient CRISPR-mediated large DNA docking and multiplexed 
            prime editing using a single baculovirus.
  JOURNAL   Nucleic Acids Res. 2022 Jul 22;50(13):7783-7799. doi: 
            10.1093/nar/gkac587.
  PUBMED    35801912
REFERENCE   2  (bases 1 to 3971)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2022-07-22- 22"; volume: "50"; issue: 
            "13"; pages: "7783-7799"
COMMENT     Sequence Label: pMMDS DEST
FEATURES             Location/Qualifiers
     source          1..3971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             967..1272
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1151..1170
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1313..1437)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    1536..1569
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      1571..1927
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             2392..3183
                     /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"
     promoter        3274..3383
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     primer_bind     complement(3475..3494)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3519..3567
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    3791..3915
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt tatcggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccagatc tggatccggc ttactaaaag ccagataaca
      661 gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg
      721 aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac
      781 agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca
      841 tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga
      901 tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct
      961 ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
     1021 gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt
     1081 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat
     1141 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa
     1201 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
     1261 tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggtcg accatagtga
     1321 ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa
     1381 tatattgata tttatatcat tttacgtttc tcgttcagct ttcttgtaca aagtggttga
     1441 taagcttaaa cgtcgaggga tcttcataag agaagaggga cagctatgac tgggagtagt
     1501 caggagagga ggaaaaaaac ccatgtgcct ggcagataac ttcgtataat gtatgctata
     1561 cgaagttatg gtacgtacta agctctcatg tttcacgtac taagctctca tgtttaacgt
     1621 actaagctct catgtttaac gaactaaacc ctcatggcta acgtactaag ctctcatggc
     1681 taacgtacta agctctcatg tttcacgtac taagctctca tgtttgaaca ataaaattaa
     1741 tataaatcag caacttaaat agcctctaag gttttaagtt ttataagaaa aaaaagaata
     1801 tataaggctt ttaaagcttt taaggtttaa cggttgtgga caacaagcca gggatgtaac
     1861 gcactgagaa gcccttagag cctctcaaag caattttgag tgacacagga acacttaacg
     1921 gctgacatgg gaattagctt cacgctgccg caagcactca gggcgcaagg gctgctaaag
     1981 gaagcggaac acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag
     2041 ctgggaggca gaataaatga tcatatcgtc aattattacc tccacgggga gagcctgagc
     2101 aaactggcct caggcatttg agaagcacac ggtcacactg cttccggtag tcaataaacc
     2161 ggtaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
     2221 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg
     2281 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     2341 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat catgagggaa
     2401 gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat
     2461 ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag
     2521 ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg
     2581 cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc
     2641 cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct
     2701 aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag
     2761 ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt
     2821 gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt
     2881 gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag
     2941 cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg
     3001 ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc
     3061 atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca
     3121 gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa
     3181 taatgtctaa caattcgttc aagccgacgg atctatgtcg ggtgcggaga aagaggtaat
     3241 gaaatggcac ctaggtatcg atactagtat acggaccttt aattcaaccc aacacaatat
     3301 attatagtta aataagaatt attatcaaat catttgtata ttaattaaaa tactatactg
     3361 taaattacat tttatttaca atcactcgac gaagacttga tcacccggga tctcgagcca
     3421 tggtgctagc agctgatgca tagcatgcgg taccgggaga tgggggaggc taactgaaac
     3481 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     3541 aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     3601 ctgtcgatac cccaccgaga ccccattggg accaatacgc ccgcgtttct tccttttccc
     3661 caccccaacc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcaa
     3721 gccctgccat agccactacg ggtacgtttc tgtaactata acggtcctaa ggtagcgagt
     3781 ttaaacatca acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg atataaatat
     3841 caatatatta aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca
     3901 tatccagtca ctatggcggc cgcattaggc cactgagcgt cagaccccgt agaaaagatc
     3961 aaaggatctt c
//
