LOCUS       sequence-469234-        8489 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of MBP-EZH2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8489)
  AUTHORS   Lai Z, Flanigan SF, Boudes M, Davidovich C
  TITLE     Modular cloning of multigene vectors for the baculovirus system and 
            yeast.
  JOURNAL   J Mol Biol. 2025 Jan 13:168943. doi: 10.1016/j.jmb.2025.168943.
  PUBMED    39814169
REFERENCE   2  (bases 1 to 8489)
  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"; journalName: "J Mol Biol.
            2025 Jan 13:168943. doi: 10.1016/j.jmb.2025.168943."
COMMENT     Sequence Label: pMB.M.E2
FEATURES             Location/Qualifiers
     source          1..8489
                     /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"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  892..1116
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1183..1716)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1905..1910,1911..1927,1928..1933))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        2481..2572
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2497..2514
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2624..3721
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      3578..3587
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3695..3718
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             3755..3778
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    6173..6307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6210..6229)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6264..6283
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6517..6682
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      6872..7327
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6959..6978)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7169..7190
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7354..7458
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7459..7527,7528..8319)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7677..7696)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct 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 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcagaccag ccgcgtaacc
      841 tggcaaaatc ggttacggtt gagtaataaa tggatgccct gcgtaagcgg gtgtgggcgg
      901 acaataaagt cttaaactga acaaaataga tctaaactat gacaataaag tcttaaacta
      961 gacagaatag ttgtaaactg aaatcagtcc agttatgctg tgaaaaagca tactggactt
     1021 ttgttatggc taaagcaaac tcttcatttt ctgaagtgca aattgcccgt cgtattaaag
     1081 aggggcgtgg ccaagggcat ggtaaagact atattcgcgg cgttgtgaca atttaccgaa
     1141 caactccgcg gccgggaagc cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg
     1201 gtcgatatca aagtgcatca cttcttcccg tatgcccaac tttgtataga gagccactgc
     1261 gggatcgtca ccgtaatctg cttgcacgta gatcacataa gcaccaagcg cgttggcctc
     1321 atgcttgagg agattgatga gcgcggtggc aatgccctgc ctccggtgct cgccggagac
     1381 tgcgagatca tagatataga tctcactacg cggctgctca aacttgggca gaacgtaagc
     1441 cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca agcgcgatga atgtcttact
     1501 acggagcaag ttcccgaggt aatcggagtc cggctgatgt tgggagtagg tggctacgtc
     1561 gccgaactca cgaccgaaaa gatcaagagc agcccgcatg gatttgactt ggtcagggcc
     1621 gagcctacat gtgcgaatga tgcccatact tgagccacct aactttgttt tagggcgact
     1681 gccctgctgc gtaacatcgt tgctgctgcg taacatcgtt gctgctccat aacatcaaac
     1741 atcgacccac ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtacaaaaa
     1801 aacagtcata acaagccatg aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca
     1861 aggttctgga ccagttgcgt gagcgcatac gctacttgca ttacagttta cgaaccgaac
     1921 aggcttatgt caactgggtt cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa
     1981 ccttgggcag cagcgaagtc gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt
     2041 cggactccac gcatcgtcag gcattggcgg ccttgctgtt cttctacggc aaggtgctgt
     2101 gcacggatct gccctggctt caggagatcg gaagacctcg gccgtcgcgg cgcttgccgg
     2161 tggtgctgac cccggatgaa gtggttcgca tcctcggttt tctggaaggc gagcatcgtt
     2221 tgttcgccca ggactctagc tatagttccc tgaattcgca tctagatggt agagccacaa
     2281 acagccggta caagcaacga tctccaggac catctgaatc atgcgcggat gacacgaact
     2341 cacgacggcg atcacagaca ttaacccaca gtacagacac tgcgacaacg tggcaattcg
     2401 tcgcaatacc gtctcactga actggccgat aattgcagac gaacgctagt ggttggctac
     2461 atactccgga atattaatag atcatggaga taattaaaat gataaccatc tcgcaaataa
     2521 ataagtattt tactgttttc gtaacagttt tgtaataaaa aaacctataa atattccgga
     2581 ttattcatac cgtcccacca tcgggcgcgg aagaatgggt accaaaactg aagaaggtaa
     2641 actggtaatc tggattaacg gcgataaagg ctataacggt ttggctgaag tcggtaagaa
     2701 attcgagaaa gataccggaa ttaaagtcac cgttgagcat ccggataaac tggaagagaa
     2761 attcccacag gttgcggcaa ctggcgatgg ccctgacatt atcttctggg cacacgaccg
     2821 ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc accccggaca aagcgttcca
     2881 ggacaagctg tatccgttta cctgggatgc cgtacgttac aacggcaagc tgattgctta
     2941 cccgatcgct gttgaagcgt tatcgctgat ttataacaaa gatttgctgc cgaacccgcc
     3001 aaaaacctgg gaagagatcc cggcgctgga taaagaactg aaagcgaaag gtaagagcgc
     3061 gctgatgttc aacctgcaag aaccgtactt cacctggccg ctgattgctg ctgacggggg
     3121 ttatgcgttc aagtatgaaa acggcaagta cgacattaaa gacgtgggcg tggataacgc
     3181 tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt aaaaacaaac acatgaatgc
     3241 agacaccgat tactccatcg cagaagctgc ctttaataaa ggcgaaacag cgatgaccat
     3301 caacggcccg tgggcatggt ccaacatcga caccagcaaa gtgaattatg gtgtaacggt
     3361 actgccgacc ttcaagggtc aaccatccaa accgttcgtt ggcgtgctga gcgcaggtat
     3421 taacgccgcc agtccgaaca aagagctggc aaaagagttc ctcgaaaact atctgctgac
     3481 tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg ggtgccgtag cgctgaagtc
     3541 ttacgaggaa gagttggcga aagatccacg tattgccgcc accatggaaa acgcccagaa
     3601 aggtgaaatc atgccgaaca tcccgcagat gtccgctttc tggtatgccg tgcgtactgc
     3661 ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa gccctgaaag acgcgcagac
     3721 tgccgcaggc accgattacg atatcccaac gaccctggaa gttctgttcc aggggccctc
     3781 cggtatgggc cagactggga agaaatctga gaagggacca gtttgttggc ggaagcgtgt
     3841 aaaatcagag tacatgcgac tgagacagct caagaggttc agacgagctg atgaagtaaa
     3901 gagtatgttt agttccaatc gtcagaaaat tttggaaaga acggaaatct taaaccaaga
     3961 atggaaacag cgaaggatac agcctgtgca catcctgact tctgtgagct cattgcgcgg
     4021 gactagggag tgttcggtga ccagtgactt ggattttcca acacaagtca tcccattaaa
     4081 gactctgaat gcagttgctt cagtacccat aatgtattct tggtcacccc tacagcagaa
     4141 ttttatggtg gaagatgaaa ctgttttaca taacattcct tatatgggag atgaagtttt
     4201 agatcaggat ggtactttca ttgaagaact aataaaaaat tatgatggga aagtacacgg
     4261 ggatagagaa tgtgggttta taaatgatga aatttttgtg gagttggtga atgcccttgg
     4321 tcaatataat gatgatgacg atgatgatga tggagatgat cctgaagaaa gagaagaaaa
     4381 gcagaaagac ctggaggatc accgagatga taaagaaagc cgcccacctc ggaaatttcc
     4441 ttctgataaa atttttgaag ccatttcctc aatgtttcca gataagggca cagcagaaga
     4501 actaaaggaa aaatataaag aactcaccga acagcagctc ccaggcgcac ttcctcctga
     4561 atgtaccccc aacatagatg gaccaaatgc taaatctgtt cagagagagc aaagcttaca
     4621 ctcctttcat acgcttttct gtaggcgatg ttttaaatat gactgcttcc tacatcgtaa
     4681 gtgcaattat tcttttcatg caacacccaa cacttataag cggaagaaca cagaaacagc
     4741 tctagacaac aaaccttgtg gaccacagtg ttaccagcat ttggagggag caaaggagtt
     4801 tgctgctgct ctcaccgctg agcggataaa gaccccacca aaacgtccag gaggccgcag
     4861 aagaggacgg cttcccaata acagtagcag gcccagcacc cccaccatta atgtgctgga
     4921 atcaaaggat acagacagtg atagggaagc agggactgaa acggggggag agaacaatga
     4981 taaagaagaa gaagagaaga aagatgaaac ttcgagctcc tctgaagcaa attctcggtg
     5041 tcaaacacca ataaagatga agccaaatat tgaacctcct gagaatgtgg agtggagtgg
     5101 tgctgaagcc tcaatgttta gagtcctcat tggcacttac tatgacaatt tctgtgccat
     5161 tgctaggtta attgggacca aaacatgtag acaggtgtat gagtttagag tcaaagaatc
     5221 tagcatcata gctccagctc ccgctgagga tgtggatact cctccaagga aaaagaagag
     5281 gaaacaccgg ttgtgggctg cacactgcag aaagatacag ctgaaaaagg acggctcctc
     5341 taaccatgtt tacaactatc aaccctgtga tcatccacgg cagccttgtg acagttcgtg
     5401 cccttgtgtg atagcacaaa atttttgtga aaagttttgt caatgtagtt cagagtgtca
     5461 aaaccgcttt ccgggatgcc gctgcaaagc acagtgcaac accaagcagt gcccgtgcta
     5521 cctggctgtc cgagagtgtg accctgacct ctgtcttact tgtggagccg ctgaccattg
     5581 ggacagtaaa aatgtgtcct gcaagaactg cagtattcag cggggctcca aaaagcatct
     5641 attgctggca ccatctgacg tggcaggctg ggggattttt atcaaagatc ctgtgcagaa
     5701 aaatgaattt atctcagaat actgtggaga gattatttct caagatgaag ctgacagaag
     5761 agggaaagtg tatgataaat acatgtgcag ctttctgttc aacttgaaca atgattttgt
     5821 ggtggatgca acccgcaagg gtaacaaaat tcgttttgca aatcattcgg taaatccaaa
     5881 ctgctatgca aaagttatga tggttaacgg tgatcacagg ataggtattt ttgccaagag
     5941 agccatccag actggcgaag agctgttttt tgattacaga tacagccagg ctgatgccct
     6001 gaagtatgtc ggcatcgaaa gagaaatgga aatccctgga tcctaatgag tcgagaagta
     6061 ctagaggatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     6121 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     6181 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6241 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6301 ctggatctga tcactgatat cgcctaggag atccggggct gccaatgaga cgacggggtc
     6361 atcacggctc atcatgcgcc aaacaaatgt gtgcaataca cgctcggatg actgcatgat
     6421 gaccgcactg actggggaca gcagatccac ctaagcctgt gagagaagca gacacccgac
     6481 agatcaaggc agttaactag tgcactgcag tacatcaacc agataagtga aatctagttc
     6541 caaactattt tgtcattttt aattttcgta ttagcttacg acgctacacc cagttcccat
     6601 ctattttgtc actcttccct aaataatcct taaaaactcc atttccaccc ctcccagttc
     6661 ccaactattt tgtccgccca cagcggggca tttttcttcc tgttatgttt ttaatcaaac
     6721 atcctgccaa ctccatgtga caaaccgtca tcttcggcta ctttttctct gtcacagaat
     6781 gaaaattttt ctgtcatctc ttcgttatta atgtttgtaa ttgactgaat atcaacgctt
     6841 atttgcagcc tgaatggcga atggggggag tacgcgccct gtagcggcgc attaagcgcg
     6901 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     6961 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     7021 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     7081 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     7141 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     7201 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     7261 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     7321 acaatttcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     7381 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     7441 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     7501 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     7561 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     7621 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     7681 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     7741 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     7801 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     7861 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     7921 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     7981 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     8041 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     8101 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     8161 gccggtgagc gtggttctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     8221 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     8281 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     8341 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     8401 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     8461 gaccccgtag aaaagatcaa aggatcttc
//
