LOCUS       sequence-406634-        3780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ENTR Vector 2 for MultiSite Gateway assembly. Encodes mKO2-hCdt1 
            under the control of CMV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3780)
  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 3780)
  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 2 CMV mKO2-hCdt1
FEATURES             Location/Qualifiers
     source          1..3780
                     /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=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        740..1043
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1044..1247
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1198..1218
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1303..1953
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAH"
     primer_bind     complement(2345..2362)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2351..2575
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    2594..2718
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2814..2833)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2825..3634
                     /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(2892..2911)
                     /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 aataaagttg gtttaaacga tatcttcata
      721 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      781 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      841 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      901 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      961 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     1021 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     1081 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     1141 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     1201 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctgga attctgcagt
     1261 cgatcgacgg taccgcgggc cctccgcggc cgcgggtccc ccatggtgag tgtgattaaa
     1321 ccagagatga agatgaggta ctacatggac ggctccgtca atgggcatga gttcacaatt
     1381 gaaggtgaag gcacaggcag accttacgag ggacatcaag agatgacact acgcgtcaca
     1441 atggccgagg gcgggccaat gcctttcgcg tttgacttag tgtcacacgt gttctgttac
     1501 ggccacagag tatttactaa atatccagaa gagataccag actatttcaa acaagcattt
     1561 cctgaaggcc tgtcatggga aaggtcgttg gagttcgaag atggtgggtc cgcttcagtc
     1621 agtgcgcata taagccttag aggaaacacc ttctaccaca aatccaaatt tactggggtt
     1681 aactttcctg ccgatggtcc tatcatgcaa aaccaaagtg ttgattggga gccatcaacc
     1741 gagaaaatta ctgccagcga cggagttctg aagggtgatg ttacgatgta cctaaaactt
     1801 gaaggaggcg gcaatcacaa atgccaaatg aagactactt acaaggcggc aaaagagatt
     1861 cttgaaatgc caggagacca ttacatcggc catcgcctcg tcaggaaaac cgaaggcaac
     1921 attactgagc aggtagaaga tgcagtagct cattactcaa tgttgccttc ccagggcgga
     1981 tatccatcac actggcggcc gctcgagccc agccccgcca ggcccgcact ccgcgccccg
     2041 tcctccgcta ccagtggcag ccgcaagcgc gcccgcccgc ccgccgcccc cggacgcgac
     2101 caggccaggc caccggcccg caggagactg cggctgtcgg tggacgaggt ttccagcccc
     2161 agtacccccg aggccccaga catcccagcc tgcccttctc cgggccagaa gataaagaaa
     2221 tccaccccgg cagcaggtca gccgccccac ctgacatccg cgcaggacca ggacaccatc
     2281 taaggggaat tcctcgagaa gctgccgctc gagtctagag ggcccgttta aacccgctga
     2341 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2401 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2461 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2521 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatgggatat
     2581 ccctaggccc gggccaactt tgtatagaaa agttgaacga gaaacgtaaa atgatataaa
     2641 tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca
     2701 acatatccag tcactatgct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     2761 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     2821 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     2881 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     2941 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     3001 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     3061 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     3121 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     3181 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     3241 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     3301 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     3361 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     3421 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     3481 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     3541 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     3601 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     3661 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     3721 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
