LOCUS       sequence-406387-        4834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ENTR Vector 1 for MultiSite Gateway assembly and production of 
            recombinant baculovirus using MultiMate assembly. Encodes EYFP 
            Tubulin under the control of CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4834)
  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 4834)
  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 EYFP Tubulin
FEATURES             Location/Qualifiers
     source          1..4834
                     /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      1283..1292
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1289..1291,1292..1294,1295..2005)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1334..1355)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1595..1614)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1942..1963
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3509..3630
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3546..3565)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3600..3619
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    3649..3772
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3868..3887)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3879..4688
                     /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(3946..3965)
                     /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 agctggttta gtgaaccgtc
     1261 agatccgcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     1321 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1381 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     1441 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg
     1501 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1561 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1621 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1681 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1741 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     1801 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1861 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga
     1921 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1981 tctcggcatg gacgagctgt acaagtccgg actcagatct cgagtgcgtg agtgcatctc
     2041 catccacgtt ggccaggctg gtgtccagat tggcaatgcc tgctgggagc tctactgcct
     2101 ggaacacggc atccagcccg atggccagat gccaagtgac aagaccattg ggggaggaga
     2161 tgactccttc aacaccttct tcagtgagac gggcgctggc aagcacgtgc cccgggctgt
     2221 gtttgtagac ttggaaccca cagtcattga tgaagttcgc actggcacct accgccagct
     2281 cttccaccct gagcagctca tcacaggcaa ggaagatgct gccaataact atgcccgagg
     2341 gcactacacc attggcaagg agatcattga ccttgtgttg gaccgaattc gcaagctggc
     2401 tgaccagtgc accggtcttc agggcttctt ggttttccac agctttggtg ggggaactgg
     2461 ttctgggttc acctccctgc tcatggaacg tctctcagtt gattatggca agaagtccaa
     2521 gctggagttc tccatttacc cagcacccca ggtttccaca gctgtagttg agccctacaa
     2581 ctccatcctc accacccaca ccaccctgga gcactctgat tgtgccttca tggtagacaa
     2641 tgaggccatc tatgacatct gtcgtagaaa cctcgatatc gagcgcccaa cctacactaa
     2701 ccttaaccgc cttattagcc agattgtgtc ctccatcact gcttccctga gatttgatgg
     2761 agccctgaat gttgacctga cagaattcca gaccaacctg gtgccctacc cccgcatcca
     2821 cttccctctg gccacatatg cccctgtcat ctctgctgag aaagcctacc atgaacagct
     2881 ttctgtagca gagatcacca atgcttgctt tgagccagcc aaccagatgg tgaaatgtga
     2941 ccctcgccat ggtaaataca tggcttgctg cctgttgtac cgtggtgacg tggttcccaa
     3001 agatgtcaat gctgccattg ccaccatcaa aaccaagcgc agcatccagt ttgtggattg
     3061 gtgccccact ggcttcaagg ttggcatcaa ctaccagcct cccactgtgg tgcctggtgg
     3121 agacctggcc aaggtacaga gagctgtgtg catgctgagc aacaccacag ccattgctga
     3181 ggcctgggct cgcctggacc acaagtttga cctgatgtat gccaagcgtg cctttgttca
     3241 ctggtacgtg ggtgagggga tggaggaagg cgagttttca gaggcccgtg aagatatggc
     3301 tgcccttgag aaggattatg aggaggttgg tgtggattct gttgaaggag agggtgagga
     3361 agaaggagag gaatactaag gatccaccgg atctagataa ctgatcataa tcagccatac
     3421 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3481 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3541 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3601 tggtttgtcc aaactcatca atgtatctta gatatcccta ggcccgggca actttgtata
     3661 ataaagttga acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc
     3721 ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta tgctggcccg
     3781 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     3841 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     3901 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     3961 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
     4021 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     4081 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     4141 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
     4201 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     4261 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     4321 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     4381 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     4441 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     4501 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     4561 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     4621 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     4681 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac
     4741 gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag
     4801 cgtcagaccc cgtagaaaag atcaaaggat cttc
//
