LOCUS       sequence-406393-        4330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ENTR Vector 4 for MultiSite Gateway assembly and production of 
            recombinant baculovirus using MultiMate assembly. Encodes mTagBFP 
            localised to Golgi organelles under the control of CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4330)
  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 4330)
  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 4 CMV GST mTagBFP
FEATURES             Location/Qualifiers
     source          1..4330
                     /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=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        712..1091
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1092..1295
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1246..1266
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1585..2280
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     polyA_signal    2310..2444
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2347..2366)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2401..2420
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    2565..3153
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2618..2638)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3036..3047)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(3169..3268)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3364..3383)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3375..4184
                     /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(3442..3461)
                     /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 aattgatgag
      661 caatgctttt ttataatgcc aactttgtat acaaaagttg gtttaaacga tgacattgat
      721 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      781 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      841 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      901 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      961 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     1021 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     1081 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     1141 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     1201 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     1261 ggcgtgtacg gtgggaggtc tatataagca gagctgaagt gccattccgc ctgacctatg
     1321 aggcttcgtg aacccttgtt gtcaggctct gctgccatgc ccggtgccag cctgcaaagg
     1381 gcatgtcgtc ttctggttgc ggtgtgtgca ttgcatctcg gggtgactct tgtgtattat
     1441 cttgcgggaa gagatctctc ccggcttcca cagttggtgg gcgtgagtac ccccttgcaa
     1501 ggcggttcaa atagcgctgc agctattgga caaagcagcg gcgaactgag gacaggcggc
     1561 gctaaagacc caccagtagc gactagcgag ctgattaagg agaacatgca catgaagctg
     1621 tacatggagg gcaccgtgga caaccatcac ttcaagtgca catccgaggg cgaaggcaag
     1681 ccctacgagg gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc
     1741 gccttcgaca tcctggctac tagcttcctc tacggcagca agaccttcat caaccacacc
     1801 cagggcatcc ccgacttctt caagcagtcc ttccctgagg gcttcacatg ggagagagtc
     1861 accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc
     1921 tgcctcatct acaacgtcaa gatcagaggg gtgaacttca catccaacgg ccctgtgatg
     1981 cagaagaaaa cactcggctg ggaggccttc accgagacgc tgtaccccgc tgacggcggc
     2041 ctggaaggca gaaacgacat ggccctgaag ctcgtgggcg ggagccatct gatcgcaaac
     2101 atcaagacca catatagatc caagaaaccc gctaagaacc tcaagatgcc tggcgtctac
     2161 tatgtggact acagactgga aagaatcaag gaggccaaca acgagaccta cgtcgagcag
     2221 cacgaggtgg cagtggccag atactgcgac ctccctagca aactggggca caagcttaat
     2281 ggatcctagt gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2341 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2401 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca ctgcttgagc
     2461 ctagaagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     2521 gagcaataac tatcataacc cctagggtat acccatctaa ttggaatcaa cctctggatt
     2581 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2641 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2701 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     2761 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     2821 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     2881 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     2941 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     3001 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3061 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3121 cgagtcggat ctccctttgg gccgcctccc cgcatcccta ggcccgggag acagctttct
     3181 tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca
     3241 ctatcagtca aaataaaatc attatttgct ggcccgtgtc tcaaaatctc tgatgttaca
     3301 ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta
     3361 atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
     3421 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     3481 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     3541 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     3601 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     3661 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     3721 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     3781 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     3841 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     3901 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     3961 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     4021 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     4081 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     4141 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     4201 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
     4261 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     4321 aaggatcttc
//
