LOCUS       sequence-469237-        7511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of MBP-RBBP4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7511)
  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 7511)
  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.R4
FEATURES             Location/Qualifiers
     source          1..7511
                     /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    5195..5329
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5232..5251)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5286..5305
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  5539..5704
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      5894..6349
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5981..6000)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6191..6212
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6376..6480
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6481..6549,6550..7341)
                     /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(6699..6718)
                     /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 tctagagcag acctagacca
     2281 catgaggctg atgtaggaca gccaccagtg gcagctaatc aacaccgcaa gatgccgatg
     2341 cacgctcata tcatcgtcgt cagcctggta gccattcgac atacggatca gggaactcga
     2401 acccagtacc gtctcagttc tgtatgggca cagacaacct aaacgctagt 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 cggtatggcc gacaaggaag cagccttcga cgacgcagtg gaagaacgag tgatcaacga
     3841 ggaatacaaa atatggaaaa agaacacccc ttttctttat gatttggtga tgacccatgc
     3901 tctggagtgg cccagcctaa ctgcccagtg gcttccagat gtaaccagac cagaagggaa
     3961 agatttcagc attcatcgac ttgtcctggg gacacacaca tcggatgaac aaaaccatct
     4021 tgttatagcc agtgtgcagc tccctaatga tgatgctcag tttgatgcgt cacactacga
     4081 cagtgagaaa ggagaatttg gaggttttgg ttcagttagt ggaaaaattg aaatagaaat
     4141 caagatcaac catgaaggag aagtaaacag ggcccgttat atgccccaga acccttgtat
     4201 catcgcaaca aagactcctt ccagtgatgt tcttgttttt gactatacaa aacatccttc
     4261 taaaccagat ccttctggag agtgcaaccc agacttgcga ctccgtggac atcagaagga
     4321 aggctatggg ctttcttgga acccaaatct cagtgggcac ttacttagtg cttcagatga
     4381 ccataccatc tgcctgtggg acatcagtgc cgttccaaag gagggaaaag tggtagatgc
     4441 gaagaccatc tttacagggc atacggcagt agtagaagat gtttcctggc atctactcca
     4501 tgagtctctg tttgggtcag ttgctgatga tcagaaactt atgatttggg atactcgttc
     4561 aaacaatact tccaaaccaa gccactcagt tgatgctcac actgctgaag tgaactgcct
     4621 ttctttcaat ccttatagtg agttcattct tgccacagga tcagctgaca agactgttgc
     4681 cttgtgggat ctgagaaatc tgaaacttaa gttgcattcc tttgagtcac ataaggatga
     4741 aatattccag gttcagtggt cacctcacaa tgagactatt ttagcttcca gtggtactga
     4801 tcgcagactg aatgtctggg atttaagtaa aattggagag gaacaatccc cagaagatgc
     4861 agaagacggg ccaccagagt tgttgtttat tcatggtggt catactgcca agatatctga
     4921 tttctcctgg aatcccaatg aaccttgggt gatttgttct gtatcagaag acaatatcat
     4981 gcaagtgtgg caaatggcag agaacattta taatgatgaa gaccctgaag gaagcgttga
     5041 tccagaagga caagggtccg gatcctaatg agtcgagaag tactagagga tcataatcag
     5101 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     5161 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     5221 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5281 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct gatcactgat
     5341 atcgcctagg agatccgggg ctgggtatga gacggcctgc actagacgaa ctaggcaaga
     5401 tgcgtccaat ccgtctaaac atggtgacaa cgctggacag atgacgtaac accgagccac
     5461 atcctgaaat cgaggcaggc taaccgaaac cgtgacaatg caaagagaca gcctgacact
     5521 agtgcactgc agtacatcaa ccagataagt gaaatctagt tccaaactat tttgtcattt
     5581 ttaattttcg tattagctta cgacgctaca cccagttccc atctattttg tcactcttcc
     5641 ctaaataatc cttaaaaact ccatttccac ccctcccagt tcccaactat tttgtccgcc
     5701 cacagcgggg catttttctt cctgttatgt ttttaatcaa acatcctgcc aactccatgt
     5761 gacaaaccgt catcttcggc tactttttct ctgtcacaga atgaaaattt ttctgtcatc
     5821 tcttcgttat taatgtttgt aattgactga atatcaacgc ttatttgcag cctgaatggc
     5881 gaatgggggg agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5941 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     6001 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     6061 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     6121 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     6181 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     6241 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     6301 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc aggtggcact
     6361 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6421 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6481 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     6541 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     6601 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6661 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     6721 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6781 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6841 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6901 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6961 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     7021 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     7081 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7141 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggttct
     7201 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     7261 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     7321 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     7381 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7441 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7501 aaaggatctt c
//
