LOCUS       sequence-406400-        4224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ENTR Vector 4 for MultiSite Gateway assembly. Encodes mTagBFP under 
            the control of a CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4224)
  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 4224)
  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 mTagBFP
FEATURES             Location/Qualifiers
     source          1..4224
                     /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"
     primer_bind     1292..1316
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1431..1700
                     /label=CMV intron
                     /note="modified intron A from human cytomegalovirus"
     CDS             1751..2449
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    2459..3047
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2512..2532)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2930..2941)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(3063..3162)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3258..3277)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3269..4078
                     /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(3336..3355)
                     /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 gagctcgttt agtgaaccgt cagatcgcct
     1321 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc
     1381 gcggccggga acggtgcatt ggaacgcgga ttccccgtgc caagagtgac gtaagtaccg
     1441 cctatagact ctataggcac acccctttgg ctcttatgca tgctatactg tttttggctt
     1501 ggggcctata cacccccgct tccttatgct ataggtgatg gtatagctta gcctataggt
     1561 gtgggttatt gaccattatt gaccactcca acggtggagg gcagtgtagt ctgagcagta
     1621 ctcgttgctg ccgcgcgcgc caccagacat aatagctgac agactaacag actgttcctt
     1681 tccatgggtc ttttctgcag tcaccgtcgt cgacggtatc gataagcttg atatcgaatt
     1741 cgccgccgcc atgagcgagc tgattaagga gaacatgcac atgaagctgt acatggaggg
     1801 caccgtggac aaccatcact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg
     1861 cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat
     1921 cctggctact agcttcctct acggcagcaa gaccttcatc aaccacaccc agggcatccc
     1981 cgacttcttc aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga
     2041 agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta
     2101 caacgtcaag atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac
     2161 actcggctgg gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag
     2221 aaacgacatg gccctgaagc tcgtgggcgg gagccatctg atcgcaaaca tcaagaccac
     2281 atatagatcc aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta
     2341 cagactggaa agaatcaagg aggccaacaa cgagacctac gtcgagcagc acgaggtggc
     2401 agtggccaga tactgcgacc tccctagcaa actggggcac aagcttaatg gatcctagaa
     2461 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2521 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2581 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2641 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2701 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2761 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2821 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     2881 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     2941 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3001 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc cctaggcccg
     3061 ggagacagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     3121 caacgaacag gtcactatca gtcaaaataa aatcattatt tgctggcccg tgtctcaaaa
     3181 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     3241 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     3301 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     3361 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     3421 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     3481 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     3541 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     3601 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     3661 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     3721 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     3781 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     3841 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     3901 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     3961 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     4021 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     4081 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     4141 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     4201 cgtagaaaag atcaaaggat cttc
//
