LOCUS       sequence-406392-        3513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ENTR Vector 3 for MultiSite Gateway assembly and production of 
            recombinant baculovirus using MultiMate assembly. Encodes 
            mitochondrially localised mCherry under the control of CMV 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3513)
  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 3513)
  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: pMMK ENTR 3 CMV mito mCherry
FEATURES             Location/Qualifiers
     source          1..3513
                     /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"
     protein_bind    605..700
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        739..1042
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1043..1246
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1197..1217
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1275..1349
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1365..2072
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1507..1525)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1775..1794
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2188..2309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2225..2244)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2279..2298
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    2328..2451
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2547..2566)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2558..3367
                     /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(2625..2644)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
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 taccggataa
      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 cgcgaaataa tgattttatt ttgactgata gtgacctgtt cgttgcaaca aattgataag
      661 caatgctttc ttataatgcc aactttgtat agaaaagttg gtttaaacga tatctcatag
      721 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      781 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      841 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      901 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      961 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     1021 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     1081 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     1141 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     1201 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     1261 tcagatccgc tagcatgtcc gtcctgacgc cgctgctgct gcggggcttg acaggctcgg
     1321 cccggcggct cccagtgccg cgcgccaaga tccattcgtt gggggtgagc aagggcgagg
     1381 aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg
     1441 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     1501 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt
     1561 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
     1621 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     1681 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
     1741 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
     1801 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
     1861 agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca
     1921 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca
     1981 cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact
     2041 ccaccggcgg catggacgag ctgtacaagt aagcgactct agatcataat cagccatacc
     2101 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2161 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2221 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2281 ggtttgtcca aactcatcaa tgtatcttag atatccctag gcccgggcaa ctttgtatac
     2341 aaaagttgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca
     2401 taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat gctggcccgt
     2461 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     2521 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     2581 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2641 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     2701 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2761 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2821 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     2881 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2941 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     3001 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3061 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3121 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3181 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3241 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3301 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     3361 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     3421 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3481 gtcagacccc gtagaaaaga tcaaaggatc ttc
//
