LOCUS       sequence-406391-        5688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ENTR Vector 1 for MultiSite Gateway assembly and production of 
            recombinant baculovirus using MultiMate assembly. Encodes mAG Ctnnb1
            under the control of CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5688)
  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 5688)
  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 1 CMV mAG beta-catenin
FEATURES             Location/Qualifiers
     source          1..5688
                     /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..704
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        737..1040
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1041..1244
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1195..1215
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1316..1325
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1322..1999
                     /codon_start=1
                     /product="humanized monomeric Azami-Green fluorescent 
                     protein"
                     /label=hmAzami-Green
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNV
                     TEGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTA
                     TSNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLL
                     LEGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLP
                     SQAK"
     CDS             1322..1999
                     /codon_start=1
                     /product="monomeric Azami-Green fluorescent protein 
                     (Karasawa et al., 2003)"
                     /label=mAzami-Green
                     /translation="MVSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNV
                     TEGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTA
                     TSNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLL
                     LEGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLP
                     SQAK"
     CDS             2042..2077
                     /codon_start=1
                     /product="nanobody epitope tag corresponding to amino acids
                     16-27 of human beta-catenin (Traenkle et al., 2015)"
                     /label=BC2 tag
                     /translation="PDRKAAVSHWQQ"
     polyA_signal    4363..4484
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4400..4419)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4454..4473
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    4503..4626
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4722..4741)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4733..5542
                     /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(4800..4819)
                     /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 cgcgcaaata atgattttat tttgactgat agtgacctgt tcgttgcaac aaattgataa
      661 gcaatgcttt tttataatgc caactttgta caaaaaagca ggctgtttaa acgatatccc
      721 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      781 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      841 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      901 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      961 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     1021 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     1081 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     1141 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     1201 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggaatt ctgcagtcga
     1261 tcgacggtac cgcgggcccg ggccactagt aacggccgcc agtgtgctgg aattcgccac
     1321 catggtgagc gtgatcaagc ccgagatgaa gatcaagctg tgcatgaggg gcaccgtgaa
     1381 cggccacaac ttcgtgatcg agggcgaggg caagggcaac ccctacgagg gcacccagat
     1441 cctggacctg aacgtgaccg agggcgcccc cctgcccttc gcctacgaca tcctgaccac
     1501 cgtgttccag tacggcaaca gggccttcac caagtacccc gccgacatcc aggactactt
     1561 caagcagacc ttccccgagg gctaccactg ggagaggagc atgacctacg aggaccaggg
     1621 catctgcacc gccaccagca acatcagcat gaggggcgac tgcttcttct acgacatcag
     1681 gttcgacggc accaacttcc cccccaacgg ccccgtgatg cagaagaaga ccctgaagtg
     1741 ggagcccagc accgagaaga tgtacgtgga ggacggcgtg ctgaagggcg acgtgaacat
     1801 gaggctgctg ctggagggcg gcggccacta caggtgcgac ttcaagacca cctacaaggc
     1861 caagaaggag gtgaggctgc ccgacgccca caagatcgac cacaggatcg agatcctgaa
     1921 gcacgacaag gactacaaca aggtgaagct gtacgagaac gccgtggcca ggtactccat
     1981 gctgcccagc caggccaagg ctactcaagc tgacctgatg gagttggaca tggccatgga
     2041 gccggacaga aaagctgctg tcagccactg gcagcagcag tcttacttgg attctggaat
     2101 ccattctggt gccaccacca cagctccttc cctgagtggc aagggcaacc ctgaggaaga
     2161 agatgttgac acctcccaag tcctttatga atgggagcaa ggcttttccc agtccttcac
     2221 gcaagagcaa gtagctgata ttgacgggca gtatgcaatg actagggctc agagggtccg
     2281 agctgccatg ttccctgaga cgctagatga gggcatgcag atcccatcca cgcagtttga
     2341 cgctgctcat cccactaatg tccagcgctt ggctgaacca tcacagatgt tgaaacatgc
     2401 agttgtcaat ttgattaact atcaggatga cgcggaactt gccacacgtg caattcctga
     2461 gctgacaaaa ctgctaaacg atgaggacca ggtggtagtt aataaagctg ctgttatggt
     2521 ccatcagctt tccaaaaagg aagcttccag acatgccatc atgcgctccc ctcagatggt
     2581 gtctgccatt gtacgcacca tgcagaatac aaatgatgta gagacagctc gttgtactgc
     2641 tgggactctg cacaaccttt ctcaccaccg cgagggcttg ctggccatct ttaagtctgg
     2701 tggcatccca gcgctggtga aaatgcttgg gtcaccagtg gattctgtac tgttctacgc
     2761 catcacgaca ctgcataatc tcctgctcca tcaggaagga gctaaaatgg cagtgcgcct
     2821 agctggtgga ctgcagaaaa tggttgcttt gctcaacaaa acaaacgtga aattcttggc
     2881 tattacaaca gactgccttc agatcttagc ttatggcaat caagagagca agctcatcat
     2941 tctggccagt ggtggacccc aagccttagt aaacataatg aggacctaca cttatgagaa
     3001 gcttctgtgg accacaagca gagtgctgaa ggtgctgtct gtctgctcta gcaacaagcc
     3061 ggccattgta gaagctggtg ggatgcaggc actggggctt catctgacag acccaagtca
     3121 gcgacttgtt caaaactgtc tttggactct cagaaacctt tcagatgcag cgactaagca
     3181 ggaagggatg gaaggcctcc ttgggactct agtgcagctt ctgggttccg atgatataaa
     3241 tgtggtcacc tgtgcagctg gaattctctc taacctcact tgcaataatt acaaaaacaa
     3301 gatgatggtg tgccaagtgg gtggcataga ggctcttgta cgcaccgtcc ttcgtgctgg
     3361 tgacagggaa gacatcactg agcctgccat ctgtgctctt cgtcatctga ccagccggca
     3421 tcaggaagcc gagatggccc agaatgccgt tcgccttcat tatggactgc ctgttgtggt
     3481 taaactcctg cacccaccat cccactggcc tctgataaag gcaactgttg gattgattcg
     3541 aaaccttgcc ctttgcccag caaatcatgc gcctttgcgg gaacagggtg ctattccacg
     3601 actagttcag ctgcttgtac gagcacatca ggacacccaa cggcgcacct ccatgggtgg
     3661 aacgcagcag cagtttgtgg agggcgtgcg catggaggag atagtagaag ggtgtactgg
     3721 agctctccac atccttgctc gggacgttca caaccggatt gtaatccgag gactcaatac
     3781 cattccattg tttgtgcagt tgctttattc tcccattgaa aatatccaaa gagtagctgc
     3841 aggggtcctc tgtgaacttg ctcaggacaa ggaggctgca gaggccattg aagctgaggg
     3901 agccacagct cccctgacag agttactcca ctccaggaat gaaggcgtgg caacatacgc
     3961 agctgctgtc ctattccgaa tgtctgagga caagccacag gattacaaga agcggctttc
     4021 agtcgagctg accagttccc tcttcaggac agagccaatg gcttggaatg agactgcaga
     4081 tcttggactg gacattggtg cccagggaga agcccttgga tatcgccagg atgatcccag
     4141 ctaccgttct tttcactctg gtggatacgg ccaggatgcc ttggggatgg accctatgat
     4201 ggagcatgag atgggtggcc accaccctgg tgctgactat ccagttgatg ggctgcctga
     4261 tctgggacac gcccaggacc tcatggatgg gctgccccca ggtgatagca atcagctggc
     4321 ctggtttgat actgacctgt aatgaatgca attgttgttg ttaacttgtt tattgcagct
     4381 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4441 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttagatatc cctaggcccg
     4501 ggcaactttg tataataaag ttgaacgaga aacgtaaaat gatataaata tcaatatatt
     4561 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc
     4621 actatgctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     4681 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     4741 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     4801 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     4861 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     4921 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     4981 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     5041 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     5101 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     5161 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     5221 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     5281 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     5341 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     5401 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     5461 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     5521 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     5581 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     5641 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
