LOCUS       sequence-316943-        3566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term MBP 3C cleavable Kan resistant bacterial expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3566)
  AUTHORS   Altmannova V, Blaha A, Astrinidis S, Reichle H, Weir JR
  TITLE     InteBac - An integrated bacterial and baculovirus expression vector 
            suite.
  JOURNAL   Protein Sci. 2020 Sep 21. doi: 10.1002/pro.3957.
  PUBMED    32955754
REFERENCE   2  (bases 1 to 3566)
  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: "Protein 
            Sci. 2020 Sep 21. doi: 10.1002/pro.3957."
COMMENT     Sequence Label: 40 pCOLI_K_MBP
FEATURES             Location/Qualifiers
     source          1..3566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(27..118)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(134..883)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(144..161)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(384..403)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1079..1098
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1079..1097
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1135..1157
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1171..1188
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1198..2298
                     /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="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     2272..2295
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2308..2331
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2486..2504)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2500..2547
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(2777..3566,1..26))
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(join(2777..3566,1..26))
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3500..3519
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gacgtttccc gttgaatatg gctcatactc ttcctttttc aatattattg aagcatttat
       61 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
      121 ggcatgcagc gctcttccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg
      181 cggcgagcgg tgtcagctca ctcaaaagcg gtaatacggt tatccacaga atcaggggat
      241 aaagccggaa agaacatgtg agcaaaaagc aaagcaccgg aagaagccaa cgccgcaggc
      301 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagccagag
      361 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      421 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      481 aagcgtggcg ctttctcata gctcacgctg ttggtatctc agttcggtgt aggtcgttcg
      541 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      601 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccat
      661 tggtaactga tttagaggac tttgtcttga agttatgcac ctgttaaggc taaactgaaa
      721 gaacagattt tggtgagtgc ggtcctccaa cccacttacc ttggttcaaa gagttggtag
      781 ctcagcgaac cttgagaaaa ccaccgttgg tagcggtggt ttttctttat ttatgagatg
      841 atgaatcaat cggtctatca agtcaacgaa cagctattcc gttactctag atttcagtgc
      901 aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct
      961 catgttagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     1021 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgcta
     1081 atacgactca ctatagggag accacaacgg tttccctccg gctccggaaa taattttgtt
     1141 taactttaag aaggagatat acatatggca caccatcacc accatcacag cagcggcatg
     1201 aaaatcgaag aaggtaaact ggtaatctgg attaacggcg ataaaggcta taacggtctc
     1261 gctgaagtcg gtaagaaatt cgagaaagat accggaatta aagtcaccgt tgagcatccg
     1321 gataaactgg aagagaaatt cccacaggtt gcggcaactg gcgatggccc tgacattatc
     1381 ttctgggcac acgaccgctt tggtggctac gctcaatctg gcctgttggc tgaaatcacc
     1441 ccggacaaag cgttccagga caagctgtat ccgtttacct gggatgccgt acgttacaac
     1501 ggcaagctga ttgcttaccc gatcgctgtt gaagcgttat cgctgattta taacaaagat
     1561 ctgctgccga acccgccaaa aacctgggaa gagatcccgg cgctggataa agaactgaaa
     1621 gcgaaaggta agagcgcgct gatgttcaac ctgcaagaac cgtacttcac ctggccgctg
     1681 attgctgctg acgggggtta tgcgttcaag tatgaaaacg gcaagtacga cattaaagac
     1741 gtgggcgtgg ataacgctgg cgcgaaagcg ggtctgacct tcctggttga cctgattaaa
     1801 aacaaacaca tgaatgcaga caccgattac tccatcgcag aagctgcctt taataaaggc
     1861 gaaacagcga tgaccatcaa cggcccgtgg gcatggtcca acatcgacac cagcaaagtg
     1921 aattatggtg taacggtact gccgaccttc aagggtcaac catccaaacc gttcgttggc
     1981 gtgctgagcg caggtattaa cgccgccagt ccgaacaaag agctggcaaa agagttcctc
     2041 gaaaactatc tgctgactga tgaaggtctg gaagcggtta ataaagacaa accgctgggt
     2101 gccgtagcgc tgaagtctta cgaggaagag ttggcgaaag atccacgtat tgccgccact
     2161 atggaaaacg cccagaaagg tgaaatcatg ccgaacatcc cgcagatgtc cgctttctgg
     2221 tatgccgtgc gtactgcggt gatcaacgcc gccagcggtc gtcagactgt cgatgaagcc
     2281 ctgaaagacg cgcagactag cagcggtctg gaagttctgt tccaggggcc cggatccgtc
     2341 gacgagctca ctagtcgcgg ccgctttcga atctagagcc tgcagtctcg acaagcttgt
     2401 cgagaagtac tagaggatgt acaacgcgtg cgcgctgatc cggctgctaa caaagcccga
     2461 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     2521 tctaaacggg tcttgagggg ttttttgctg aaacctcagg catttgagaa gcacacggtc
     2581 acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa
     2641 cgaccctgcc ctgaaccgac gacaagctga cgaccgggtc tccgcaagtg gcacttttcg
     2701 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2761 gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     2821 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     2881 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     2941 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     3001 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     3061 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     3121 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg gtcgctgtta aaaggacaat
     3181 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     3241 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     3301 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     3361 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     3421 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     3481 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     3541 tggaatttaa tcgcggccta gagcaa
//
