LOCUS       sequence-406402-        4701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ENTR Vector 2 for MultiSite Gateway assembly and production of 
            recombinant baculovirus using MultiMate assembly. Encodes mTFP1 
            Actin under the control of CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4701)
  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 4701)
  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 mTFP1 Actin
FEATURES             Location/Qualifiers
     source          1..4701
                     /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"
     primer_bind     1292..1311
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1292..1310
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1372..2079
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     primer_bind     complement(3266..3283)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3272..3496
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3515..3639
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3735..3754)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3746..4555
                     /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(3813..3832)
                     /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 cagagctctc tggctaacta
     1261 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
     1321 ctggctagcg tttaaactta agcttggtac cgagctcaaa ggagatatac catggtgagc
     1381 aagggcgagg agaccacaat gggcgtaatc aagcccgaca tgaagatcaa gctgaagatg
     1441 gagggcaacg tgaatggcca cgccttcgtg atcgagggcg agggcgaggg caagccctac
     1501 gacggcacca acaccatcaa cctggaggtg aaggagggag cccccctgcc cttctcctac
     1561 gacattctga ccaccgcgtt cgcctacggc aacagggcct tcaccaagta ccccgacgac
     1621 atccccaact acttcaagca gtccttcccc gagggctact cttgggagcg caccatgacc
     1681 ttcgaggaca agggcatcgt gaaggtgaag tccgacatct ccatggagga ggactccttc
     1741 atctacgaga tacacctcaa gggcgagaac ttccccccca acggccccgt gatgcagaag
     1801 aagaccaccg gctgggacgc ctccaccgag aggatgtacg tgcgcgacgg cgtgctgaag
     1861 ggcgacgtca agcacaagct gctgctggag ggcggcggcc accaccgcgt tgacttcaag
     1921 accatctaca gggccaagaa ggcggtgaag ctgcccgact atcactttgt ggaccaccgc
     1981 atcgagatcc tgaaccacga caaggactac aacaaggtga ccgtttacga gagcgccgtg
     2041 gcccgcaact ccaccgacgg catggacgag ctgtacaagg gatcaggatc aggatcaatg
     2101 gatgatgata tcgccgcgct cgtcgtcgac aacggctccg gcatgtgcaa ggccggcttc
     2161 gcgggcgacg atgccccccg ggccgtcttc ccctccatcg tggggcgccc caggcaccag
     2221 ggcgtgatgg tgggcatggg tcagaaggat tcctatgtgg gcgacgaggc ccagagcaag
     2281 agaggcatcc tcaccctgaa gtaccccatc gagcacggca tcgtcaccaa ctgggacgac
     2341 atggagaaaa tctggcacca caccttctac aatgagctgc gtgtggctcc cgaggagcac
     2401 cccgtgctgc tgaccgaggc ccccctgaac cccaaggcca accgcgagaa gatgacccag
     2461 atcatgtttg agaccttcaa caccccagcc atgtacgttg ctatccaggc tgtgctatcc
     2521 ctgtacgcct ctggccgtac cactggcatc gtgatggact ccggtgacgg ggtcacccac
     2581 actgtgccca tctacgaggg gtatgccctc ccccatgcca tcctgcgtct ggacctggct
     2641 ggccgggacc tgactgacta cctcatgaag atcctcaccg agcgcggcta cagcttcacc
     2701 accacggccg agcgggaaat cgtgcgtgac attaaggaga agctgtgcta cgtcgccctg
     2761 gacttcgagc aagagatggc cacggctgct tccagctcct ccctggagaa gagctacgag
     2821 ctgcctgacg gccaggtcat caccattggc aatgagcggt tccgctgccc tgaggcactc
     2881 ttccagcctt ccttcctggg catggagtcc tgtggcatcc acgaaactac cttcaactcc
     2941 atcatgaagt gtgacgtgga catccgcaaa gacctgtacg ccaacacagt gctgtctggc
     3001 ggcaccacca tgtaccctgg cattgccgac aggatgcaga aggagatcac tgccctggca
     3061 cccagcacaa tgaagatcaa gatcattgct cctcctgagc gcaagtactc cgtgtggatc
     3121 ggcggctcca tcctggcctc gctgtccacc ttccagcaga tgtggatcag caagcaggag
     3181 tatgacgagt ccggcccctc catcgtccac cgcaaatgct tctgaccgct cgagtctaga
     3241 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3301 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3361 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3421 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3481 gcggtgggct ctatgggata tccctaggcc cgggccaact ttgtatagaa aagttgaacg
     3541 agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata aaaaacagac
     3601 tacataatac tgtaaaacac aacatatcca gtcactatgc tggcccgtgt ctcaaaatct
     3661 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     3721 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     3781 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     3841 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     3901 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     3961 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     4021 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     4081 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     4141 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     4201 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     4261 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     4321 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     4381 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     4441 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     4501 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     4561 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     4621 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     4681 agaaaagatc aaaggatctt c
//
