LOCUS       sequence-406403-        6510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DEST vector for MultiSite Gateway assembly and production of 
            recombinant baculovirus using MultiMate assembly. Encodes H2B 
            iRFP713 under the control of CMV promoter. CRE-Acceptor in MultiBac 
            system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6510)
  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 6510)
  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: pMMACE DEST CMV H2B iRFP
FEATURES             Location/Qualifiers
     source          1..6510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..125)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        174..553
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        554..757
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     708..728
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             815..1192
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1205..1214
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2412..2546
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2449..2468)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2503..2522
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2667..2832
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3022..3610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3511..3530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  3745..3968
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4035..4568)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4757..4762,4763..4779,4780..4785))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(4856..4889)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4931..5055
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(5080..5085,5086..5103,5104..5110)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             5164..5823
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5230..5249)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             6165..6470
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     6349..6368
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 catagtgact ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc
       61 taatttaata tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa
      121 gtggttgata agcttaaaca ctagtatcga tcgcgatgta cgggccagat atagacattg
      181 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      241 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      301 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      361 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      421 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      481 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      541 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      601 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      661 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      721 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
      781 tgcttactgg cttatgtacc gcgggcccgc cgccatgcca gagccagcga agtctgctcc
      841 cgccccgaaa aagggctcca agaaggcggt gactaaggcg cagaagaaag gcggcaagaa
      901 gcgcaagcgc agccgcaagg agagctattc catctatgtg tacaaggttc tgaagcaggt
      961 ccaccctgac accggcattt cgtccaaggc catgggcatc atgaattcgt ttgtgaacga
     1021 cattttcgag cgcatcgcag gtgaggcttc ccgcctggcg cattacaaca agcgctcgac
     1081 catcacctcc agggagatcc agacggccgt gcgcctgctg ctgcctgggg agttggccaa
     1141 gcacgccgtg tccgagggta ctaaggccat caccaagtac accagcgcta aggatccacc
     1201 ggtcgccacc atggccgaag ggagcgtggc aagacagccc gatctgctga catgcgacga
     1261 cgaaccaatc cacattccag gagctatcca gcctcatgga ctgctgctgg ctctggccgc
     1321 tgacatgact atcgtggcag gctctgataa cctgccagag ctgaccgggc tggctatcgg
     1381 agcactgatt ggccggtccg cagccgacgt gttcgattct gagactcaca atagactgac
     1441 catcgccctg gctgaacctg gggctgcagt gggagcccca attaccgtcg gcttcacaat
     1501 gcgaaaggac gctggcttta tcgggtcctg gcaccgccat gatcagctga tttttctgga
     1561 gctggaacca cctcagagag acgtggcaga gccacaggct ttctttcgga gaacaaacag
     1621 cgccatcagg cgcctgcagg ccgctgagac tctggaatcc gcttgcgcag ccgctgcaca
     1681 ggaagtgagg aagatcaccg ggttcgatcg cgtcatgatc tacagattcg cctcagactt
     1741 tagcggagaa gtgatcgcag aagatcgctg tgccgaggtc gaaagtaaac tgggcctgca
     1801 ttacccagcc tcaaccgtgc ctgcacaggc acgacggctg tatacaatca accccgtgcg
     1861 gatcattccc gacattaatt acagaccagt gcccgtcaca cctgacctga acccagtgac
     1921 tgggaggccc atcgatctga gcttcgccat tctgcgcagc gtgtcccctg tccacctgga
     1981 gtttatgagg aatatcggaa tgcatggcac catgtctatc agtattctga gaggcgaaag
     2041 gctgtggggg ctgatcgtgt gccaccatcg gacaccttac tatgtggacc tggacggcag
     2101 gcaggcttgt gaactggtgg ctcaggtgct ggcttggcag attggcgtga tgaaggagga
     2161 ataataaggg gagacccaag ctggctagtg gatcccggtc cgaagcgcgc ggaattcaaa
     2221 ggcctacgtc gacgagctca cttgtcgcgg ccgctttcga atctagagcc tgcagtctcg
     2281 acaagcttgt cgagaagtac tagaggatca taatcagcca taccacattt gtagaggttt
     2341 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     2401 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2461 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     2521 tcaatgtatc ttatcatgtc tggatctgat cactgcttga gcctagaaga tccggctgct
     2581 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actatcataa
     2641 cccctagggt atacccatct aattggaacc agataagtga aatctagttc caaactattt
     2701 tgtcattttt aattttcgta ttagcttacg acgctacacc cagttcccat ctattttgtc
     2761 actcttccct aaataatcct taaaaactcc atttccaccc ctcccagttc ccaactattt
     2821 tgtccgccca caaccggttg acttgggtca actgtcagac caagtttact catatatact
     2881 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2941 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3001 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3061 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3121 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3181 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3241 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3301 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3361 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3421 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3481 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3541 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3601 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3661 tgctcacatg ttctttcctg cgttatcccc tgattgactt gggtcgctct tcctgtggat
     3721 gcgcagatgc cctgcgtaag cgggtgtggg cggacaataa agtcttaaac tgaacaaaat
     3781 agatctaaac tatgacaata aagtcttaaa ctagacagaa tagttgtaaa ctgaaatcag
     3841 tccagttatg ctgtgaaaaa gcatactgga cttttgttat ggctaaagca aactcttcat
     3901 tttctgaagt gcaaattgcc cgtcgtatta aagaggggcg tggccaaggg catgtaaaga
     3961 ctatattcgc ggcgttgtga caatttaccg aacaactccg cggccgggaa gccgatctcg
     4021 gcttgaacga attgttaggt ggcggtactt gggtcgatat caaagtgcat cacttcttcc
     4081 cgtatgccca actttgtata gagagccact gcgggatcgt caccgtaatc tgcttgcacg
     4141 tagatcacat aagcaccaag cgcgttggcc tcatgcttga ggagattgat gagcgcggtg
     4201 gcaatgccct gcctccggtg ctcgccggag actgcgagat catagatata gatctcacta
     4261 cgcggctgct caaacttggg cagaacgtaa gccgcgagag cgccaacaac cgcttcttgg
     4321 tcgaaggcag caagcgcgat gaatgtctta ctacggagca agttcccgag gtaatcggag
     4381 tccggctgat gttgggagta ggtggctacg tctccgaact cacgaccgaa aagatcaaga
     4441 gcagcccgca tggatttgac ttggtcaggg ccgagcctac atgtgcgaat gatgcccata
     4501 cttgagccac ctaactttgt tttagggcga ctgccctgct gcgtaacatc gttgctgctg
     4561 cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac gcgcttgctg
     4621 cttggatgcc cgaggcatag actgtacaaa aaaacagtca taacaagcca tgaaaaccgc
     4681 cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc gtgagcgcat
     4741 acgctacttg cattacagtt tacgaaccga acaggcttat gtcaactggg ttcgtgcctt
     4801 catccgtttc cacggtgtgc gtcacccggc aaccttgggc agcagcgaag tcgccataac
     4861 ttcgtatagc atacattata cgaagttatc tgtaactata acggtcctaa ggtagcgagt
     4921 ttaaacatca acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg atataaatat
     4981 caatatatta aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca
     5041 tatccagtca ctatggcggc cgcattaggc accccaggct ttacacttta tgcttccggc
     5101 tcgtataatg tgtggatttt gagttaggat ccgtcgagat tttcaggagc taaggaagct
     5161 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
     5221 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
     5281 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt
     5341 attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac
     5401 ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact
     5461 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata
     5521 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt
     5581 gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac
     5641 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa
     5701 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc
     5761 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg
     5821 taaagatctg gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat
     5881 ttttgcggta taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg
     5941 ctatgaagca gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat
     6001 atatgatgtc aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct
     6061 gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat
     6121 tgaaatgaac ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt
     6181 acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg
     6241 acacgcccgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag
     6301 tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca
     6361 ccgatatggc cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc
     6421 gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct
     6481 cccttataca cagccagtct gcaggtcgac
//
