LOCUS       sequence-469239-        7559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of MBP-EED.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7559)
  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 7559)
  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.ED
FEATURES             Location/Qualifiers
     source          1..7559
                     /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    5243..5377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5280..5299)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5334..5353
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  5587..5752
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      5942..6397
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6029..6048)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6239..6260
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6424..6528
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6529..6597,6598..7389)
                     /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(6747..6766)
                     /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 tctagaataa atcgcagcca
     2281 agtgagtgaa tagatgacgc accacggtca gacacggcca catcgtatct cacaggagca
     2341 agcgcgatag gagcactcac acatagtacg gtgatccgct gactcctttg cccaaataag
     2401 acgtgagccc gtctcagatg cacacactgg cttaagatga 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 cggtatgtcc gagagggaag tgtcgactgc gccggcggga acagacatgc ctgcggccaa
     3841 gaagcagaag ctgagcagtg acgagaacag caatccagac ctctctggag atgagaatga
     3901 tgacgctgtc agtatagaaa gtggtacaaa cactgaacgc cctgatacac ctacaaacac
     3961 gccaaatgca cctggaagga aaagttgggg aaagggaaaa tggaagtcaa agaaatgcaa
     4021 atattctttc aaatgtgtaa atagtctcaa ggaagatcat aaccaaccat tgtttggagt
     4081 tcagtttaac tggcacagta aagaaggaga tccattagtg tttgcaactg taggaagcaa
     4141 cagagttacc ttgtatgaat gtcattcaca aggagaaatc cggttgttgc aatcttacgt
     4201 ggatgctgat gctgatgaaa acttttacac ttgtgcatgg acctatgata gcaatacgag
     4261 ccatcctctg ctggctgtag ctggatctag aggcataatt aggataataa atcctataac
     4321 aatgcagtgt ataaagcact atgttggcca tggaaatgct atcaatgagc tgaaattcca
     4381 tccaagagat ccaaatcttc tcctgtcagt aagtaaagat catgctttac gattatggaa
     4441 tatccagacg gacactctgg tggcaatatt tggaggcgta gaagggcaca gagatgaagt
     4501 tctaagtgct gattatgatc ttttgggtga aaaaataatg tcctgtggta tggatcattc
     4561 tcttaaactt tggaggatca attcaaagag aatgatgaat gcaattaagg aatcttatga
     4621 ttataatcca aataaaacta acaggccatt tatttctcag aaaatccatt ttcctgattt
     4681 ttctaccaga gacatacata ggaattatgt tgattgtgtg cgatggttag gcgatttgat
     4741 actttctaag tcttgtgaaa atgccattgt gtgctggaaa cctggcaaga tggaagatga
     4801 tatagataaa attaaaccca gtgaatctaa tgtgactatt cttgggcgat ttgattacag
     4861 ccagtgtgac atttggtaca tgaggttttc tatggatttc tggcaaaaga tgcttgcatt
     4921 gggcaatcaa gttggcaaac tttatgtttg ggatttagaa gtagaagatc ctcataaagc
     4981 caaatgtaca acactgactc atcataaatg tggtgctgct attcgacaaa ccagttttag
     5041 cagggatagc agcattctta tagctgtttg tgatgatgcc agtatttggc gctgggatcg
     5101 acttcgagga tcctaatgag tcgagaagta ctagaggatc ataatcagcc ataccacatt
     5161 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     5221 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     5281 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5341 gtccaaactc atcaatgtat cttatcatgt ctggatctga tcactgatat cgcctaggag
     5401 atccggggct ggttctgaga cggcagacct agaccacatg aggctgatgt aggacagcca
     5461 ccagtggcag ctaatcaaca ccgcaagatg ccgatgcacg ctcatatcat cgtcgtcagc
     5521 ctggtagcca ttcgacatac ggatcaggga actcgaaccc agtacactag tgcactgcag
     5581 tacatcaacc agataagtga aatctagttc caaactattt tgtcattttt aattttcgta
     5641 ttagcttacg acgctacacc cagttcccat ctattttgtc actcttccct aaataatcct
     5701 taaaaactcc atttccaccc ctcccagttc ccaactattt tgtccgccca cagcggggca
     5761 tttttcttcc tgttatgttt ttaatcaaac atcctgccaa ctccatgtga caaaccgtca
     5821 tcttcggcta ctttttctct gtcacagaat gaaaattttt ctgtcatctc ttcgttatta
     5881 atgtttgtaa ttgactgaat atcaacgctt atttgcagcc tgaatggcga atggggggag
     5941 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6001 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6061 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6121 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6181 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6241 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     6301 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     6361 aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt tcggggaaat
     6421 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     6481 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     6541 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     6601 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     6661 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     6721 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     6781 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     6841 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6901 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6961 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     7021 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     7081 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     7141 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     7201 gctggctggt ttattgctga taaatctgga gccggtgagc gtggttctcg cggtatcatt
     7261 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     7321 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     7381 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     7441 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     7501 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
