LOCUS       sequence-469228-        8453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of MBP-SUZ12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8453)
  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 8453)
  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.S12
FEATURES             Location/Qualifiers
     source          1..8453
                     /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    6137..6271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6174..6193)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6228..6247
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6481..6646
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      6836..7291
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6923..6942)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7133..7154
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7318..7422
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7423..7491,7492..8283)
                     /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(7641..7660)
                     /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 tctagaacgg ggtcatcacg
     2281 gctcatcatg cgccaaacaa atgtgtgcaa tacacgctcg gatgactgca tgatgaccgc
     2341 actgactggg gacagcagat ccacctaagc ctgtgagaga agcagacacc cgacagatca
     2401 aggcagttac gtctcaccaa accagatgtc aacacagcta caacgctagt 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 cggtatggcg cctcagaagc acggcggtgg gggagggggc ggctcggggc ccagcgcggg
     3841 gtccggggga ggcggcttcg ggggttcggc ggcggtggcg gcggcgacgg cttcgggcgg
     3901 caaatccggc ggcgggagct gtggaggggg tggcagttac tcggcctcct cctcctcctc
     3961 cgcggcggca gcggcggggg ctgcggtgtt accggtgaag aagccgaaaa tggagcacgt
     4021 ccaggctgac cacgagcttt tcctccaggc ctttgagaag ccaacacaaa tctatagatt
     4081 tcttcgaact cggaatctca tagcaccaat atttttgcac agaactctta cttacatgtc
     4141 tcatcgaaac tccagaacaa acatcaaaag gaaaacattt aaagttgatg atatgttatc
     4201 aaaagtagag aaaatgaaag gagagcaaga atctcatagc ttgtcagctc atttgcagct
     4261 tacgtttact ggtttcttcc acaaaaatga taagccatca ccaaactcag aaaatgaaca
     4321 aaattctgtt accctggaag tcctgcttgt gaaagtttgc cacaaaaaaa gaaaggatgt
     4381 aagttgtcca ataaggcaag ttcccacagg taaaaagcag gtgcctttga atcctgacct
     4441 caatcaaaca aaacccggaa atttcccgtc ccttgcagtt tccagtaatg aatttgaacc
     4501 tagtaacagc catatggtga agtcttactc gttgctattt agagtgactc gtccaggaag
     4561 aagagagttt aatggaatga ttaatggaga aaccaatgaa aatattgatg tcaatgaaga
     4621 gcttccagcc agaagaaaac gaaatcgtga ggatggggaa aagacatttg ttgcacaaat
     4681 gacagtattt gataaaaaca ggcgcttaca gcttttagat ggggaatatg aagtagccat
     4741 gcaggaaatg gaagaatgtc caataagcaa gaaaagagca acatgggaga ctattcttga
     4801 tgggaagagg ctgcctccat tcgaaacatt ttctcaggga cctacgttgc agttcactct
     4861 tcgttggaca ggagaaacca atgataaatc tacggctcct attgccaaac ctcttgccac
     4921 tagaaattca gagagtctcc atcaggaaaa caagcctggt tcagttaaac ctactcaaac
     4981 tattgctgtt aaagaatcat tgactacaga cctacaaaca agaaaagaaa aggatactcc
     5041 aaatgaaaac cgacaaaaat taagaatatt ttatcagttt ctctataaca acaatacaag
     5101 gcaacaaact gaagcaagag atgacctgca ttgcccttgg tgtactctga actgccgcaa
     5161 actttatagt ttactcaagc atcttaaact ctgccatagc agatttatct tcaactatgt
     5221 ttatcatcca aaaggtgcta ggatagatgt ttctatcaat gagtgttatg atggctccta
     5281 tgcaggaaat cctcaggata ttcatcgcca acctggattt gcttttagtc gcaacggacc
     5341 agttaagaga acacctatca cacatattct tgtgtgcagg ccaaaacgaa caaaagcaag
     5401 catgtctgaa tttcttgaat ctgaagatgg ggaagtagaa cagcaaagaa catatagtag
     5461 tggccacaat cgtctgtatt tccatagtga tacctgctta cctctccgtc cacaagaaat
     5521 ggaagtagat agtgaagatg aaaaagatcc tgaatggcta agagaaaaaa ccattacaca
     5581 aattgaagag ttttctgatg ttaatgaagg agagaaagaa gtgatgaaac tctggaatct
     5641 ccatgtcatg aagcatgggt ttattgctga caatcaaatg aatcatgcct gtatgctgtt
     5701 tgtagaaaat tatggacaga aaataattaa gaagaattta tgtcgaaact tcatgcttca
     5761 tctagtcagc atgcatgact ttaatcttat tagcataatg tcaatagata aagctgttac
     5821 caagctccgt gaaatgcagc aaaaattaga aaagggggaa tctgcttccc ctgcaaacga
     5881 agaaataact gaagaacaaa atgggacagc aaatggattt agtgaaatta actcaaaaga
     5941 gaaagctttg gaaacagata gtgtctcagg ggtttcaaaa cagagcaaaa aacaaaaact
     6001 cggatcctaa tgagtcgaga agtactagag gatcataatc agccatacca catttgtaga
     6061 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     6121 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6181 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6241 actcatcaat gtatcttatc atgtctggat ctgatcactg atatcgccta ggagatccgg
     6301 ggctggatgt gagacgataa atcgcagcca agtgagtgaa tagatgacgc accacggtca
     6361 gacacggcca catcgtatct cacaggagca agcgcgatag gagcactcac acatagtacg
     6421 gtgatccgct gactcctttg cccaaataag acgtgagcca ctagtgcact gcagtacatc
     6481 aaccagataa gtgaaatcta gttccaaact attttgtcat ttttaatttt cgtattagct
     6541 tacgacgcta cacccagttc ccatctattt tgtcactctt ccctaaataa tccttaaaaa
     6601 ctccatttcc acccctccca gttcccaact attttgtccg cccacagcgg ggcatttttc
     6661 ttcctgttat gtttttaatc aaacatcctg ccaactccat gtgacaaacc gtcatcttcg
     6721 gctacttttt ctctgtcaca gaatgaaaat ttttctgtca tctcttcgtt attaatgttt
     6781 gtaattgact gaatatcaac gcttatttgc agcctgaatg gcgaatgggg ggagtacgcg
     6841 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6901 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6961 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     7021 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     7081 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     7141 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     7201 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     7261 aattttaaca aaatattaac gtttacaatt tcaggtggca cttttcgggg aaatgtgcgc
     7321 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     7381 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     7441 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     7501 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     7561 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     7621 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     7681 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     7741 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     7801 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     7861 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     7921 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     7981 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     8041 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     8101 tggtttattg ctgataaatc tggagccggt gagcgtggtt ctcgcggtat cattgcagca
     8161 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     8221 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     8281 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     8341 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     8401 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttc
//
