LOCUS       sequence-310327-        3518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term MBP 3C cleavable Spec resistant bacterial expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3518)
  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 3518)
  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: 30 pCOLI_S_MBP
FEATURES             Location/Qualifiers
     source          1..3518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     707..726
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        707..725
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             763..785
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             799..816
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             826..1926
                     /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     1900..1923
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1936..1959
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2114..2132)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2128..2175
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2346..3137)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(3138..3229)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(3277..3518,1..497))
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
ORIGIN
        1 ctccctcttg cgctctcctg ttccgaccct gccgtttacc ggatacctgt tccgcctttc
       61 tcccttacgg gaagtgtggc gctttctcat agctcacaca ctggtatctc ggctcggtgt
      121 aggtcgttcg ctccaagctg ggctgtaagc aagaactccc cgttcagccc gactgctgcg
      181 ccttatccgg taactgttca cttgagtcca acccggaaaa gcacggtaaa acgccactgg
      241 cagcagccat tggtaactgg gagttcgcag aggatttgtt tagctaaaca cgcggttgct
      301 cttgaagtgt gcgccaaagt ccggctacac tggaaggaca gatttggttg ctgtgctctg
      361 cgaaagccag ttaccacggt taagcagttc cccaactgac ttaaccttcg atcaaaccac
      421 ctccccaggt ggttttttcg tttacagggc aaaagattac gcgcagaaaa aaaggatctc
      481 aagaagatcc tttgatcttt tctactgaac cgctctagat ttcagtgcaa tttatctctt
      541 caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcat
      601 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
      661 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctaat acgactcact
      721 atagggagac cacaacggtt tccctccggc tccggaaata attttgttta actttaagaa
      781 ggagatatac atatggcaca ccatcaccac catcacagca gcggcatgaa aatcgaagaa
      841 ggtaaactgg taatctggat taacggcgat aaaggctata acggtctcgc tgaagtcggt
      901 aagaaattcg agaaagatac cggaattaaa gtcaccgttg agcatccgga taaactggaa
      961 gagaaattcc cacaggttgc ggcaactggc gatggccctg acattatctt ctgggcacac
     1021 gaccgctttg gtggctacgc tcaatctggc ctgttggctg aaatcacccc ggacaaagcg
     1081 ttccaggaca agctgtatcc gtttacctgg gatgccgtac gttacaacgg caagctgatt
     1141 gcttacccga tcgctgttga agcgttatcg ctgatttata acaaagatct gctgccgaac
     1201 ccgccaaaaa cctgggaaga gatcccggcg ctggataaag aactgaaagc gaaaggtaag
     1261 agcgcgctga tgttcaacct gcaagaaccg tacttcacct ggccgctgat tgctgctgac
     1321 gggggttatg cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat
     1381 aacgctggcg cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa caaacacatg
     1441 aatgcagaca ccgattactc catcgcagaa gctgccttta ataaaggcga aacagcgatg
     1501 accatcaacg gcccgtgggc atggtccaac atcgacacca gcaaagtgaa ttatggtgta
     1561 acggtactgc cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca
     1621 ggtattaacg ccgccagtcc gaacaaagag ctggcaaaag agttcctcga aaactatctg
     1681 ctgactgatg aaggtctgga agcggttaat aaagacaaac cgctgggtgc cgtagcgctg
     1741 aagtcttacg aggaagagtt ggcgaaagat ccacgtattg ccgccactat ggaaaacgcc
     1801 cagaaaggtg aaatcatgcc gaacatcccg cagatgtccg ctttctggta tgccgtgcgt
     1861 actgcggtga tcaacgccgc cagcggtcgt cagactgtcg atgaagccct gaaagacgcg
     1921 cagactagca gcggtctgga agttctgttc caggggcccg gatccgtcga cgagctcact
     1981 agtcgcggcc gctttcgaat ctagagcctg cagtctcgac aagcttgtcg agaagtacta
     2041 gaggatgtac aacgcgtgcg cgctgatccg gctgctaaca aagcccgaaa ggaagctgag
     2101 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     2161 ttgaggggtt ttttgctgaa acctcaggca tttgagaagc acacggtcac actgcttccg
     2221 gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct
     2281 gaaccgacga ccgggtcatc gtggccggat cttgcggccc ctcggcttga acgaattgtt
     2341 agacattatt tgccgactac cttggtgatc tcgcctttca cgtagtggac aaattcttcc
     2401 aactgatctg cgcgcgaggc caagcgatct tcttcttgtc caagataagc ctgtctagct
     2461 tcaagtatga cgggctgata ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca
     2521 tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact
     2581 acatttcgct catcgccagc ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt
     2641 agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga gttcctccgc cgctggacct
     2701 accaaggcaa cgctatgttc tcttgctttt gtcagcaaga tagccagatc aatgtcgatc
     2761 gtggctggct cgaagatacc tgcaagaatg tcattgcgct gccattctcc aaattgcagt
     2821 tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct
     2881 acagcgcgga gaatctcgct ctctccaggg gaagccgaag tttccaaaag gtcgttgatc
     2941 aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg taaccagcaa atcaatatca
     3001 ctgtgtggct tcaggccgcc atccactgcg gagccgtaca aatgtacggc cagcaacgtc
     3061 ggttcgagat ggcgctcgat gacgccaact acctctgata gttgagtcga tacttcggcg
     3121 atcaccgctt ccctcatact cttccttttt caatattatt gaagcattta tcagggttat
     3181 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat agctagctca
     3241 ctcggtcgct acgctccggg cgtgagactg cggcgggcgc tgcggacaca tacaaagtta
     3301 cccacagatt ccgtggataa gcaggggact aacatgtgag gcaaaacagc agggccgcgc
     3361 cggtggcgtt tttccatagg ctccgccctc ctgccagagt tcacataaac agacgctttt
     3421 ccggtgcatc tgtgggagcc gtgaggctca accatgaatc tgacagtacg ggcgaaaccc
     3481 gacaggactt aaagatcccc accgtttccg gcgggtcg
//
