LOCUS       sequence-319347-        3519 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 3519)
  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 3519)
  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..3519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     708..727
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        708..726
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             764..786
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             800..817
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             827..1927
                     /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     1901..1924
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1937..1960
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2115..2133)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2129..2176
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2347..3138)
                     /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(3139..3230)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(3278..3519,1..498))
                     /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 taagncagtt ssssaactgn cttaaccttc gatcaaacca
      421 cctccccagg tggttttttc gtttacaggg caaaagatta cgcgcagaaa aaaaggatct
      481 caagaagatc ctttgatctt ttctactgaa ccgctctaga tttcagtgca atttatctct
      541 tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc atgttagtca
      601 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
      661 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctaa tacgactcac
      721 tatagggaga ccacaacggt ttccctccgg ctccggaaat aattttgttt aactttaaga
      781 aggagatata catatggcac accatcacca ccatcacagc agcggcatga aaatcgaaga
      841 aggtaaactg gtaatctgga ttaacggcga taaaggctat aacggtctcg ctgaagtcgg
      901 taagaaattc gagaaagata ccggaattaa agtcaccgtt gagcatccgg ataaactgga
      961 agagaaattc ccacaggttg cggcaactgg cgatggccct gacattatct tctgggcaca
     1021 cgaccgcttt ggtggctacg ctcaatctgg cctgttggct gaaatcaccc cggacaaagc
     1081 gttccaggac aagctgtatc cgtttacctg ggatgccgta cgttacaacg gcaagctgat
     1141 tgcttacccg atcgctgttg aagcgttatc gctgatttat aacaaagatc tgctgccgaa
     1201 cccgccaaaa acctgggaag agatcccggc gctggataaa gaactgaaag cgaaaggtaa
     1261 gagcgcgctg atgttcaacc tgcaagaacc gtacttcacc tggccgctga ttgctgctga
     1321 cgggggttat gcgttcaagt atgaaaacgg caagtacgac attaaagacg tgggcgtgga
     1381 taacgctggc gcgaaagcgg gtctgacctt cctggttgac ctgattaaaa acaaacacat
     1441 gaatgcagac accgattact ccatcgcaga agctgccttt aataaaggcg aaacagcgat
     1501 gaccatcaac ggcccgtggg catggtccaa catcgacacc agcaaagtga attatggtgt
     1561 aacggtactg ccgaccttca agggtcaacc atccaaaccg ttcgttggcg tgctgagcgc
     1621 aggtattaac gccgccagtc cgaacaaaga gctggcaaaa gagttcctcg aaaactatct
     1681 gctgactgat gaaggtctgg aagcggttaa taaagacaaa ccgctgggtg ccgtagcgct
     1741 gaagtcttac gaggaagagt tggcgaaaga tccacgtatt gccgccacta tggaaaacgc
     1801 ccagaaaggt gaaatcatgc cgaacatccc gcagatgtcc gctttctggt atgccgtgcg
     1861 tactgcggtg atcaacgccg ccagcggtcg tcagactgtc gatgaagccc tgaaagacgc
     1921 gcagactagc agcggtctgg aagttctgtt ccaggggccc ggatccgtcg acgagctcac
     1981 tagtcgcggc cgctttcgaa tctagagcct gcagtctcga caagcttgtc gagaagtact
     2041 agaggatgta caacgcgtgc gcgctgatcc ggctgctaac aaagcccgaa aggaagctga
     2101 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     2161 cttgaggggt tttttgctga aacctcaggc atttgagaag cacacggtca cactgcttcc
     2221 ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac gaccctgccc
     2281 tgaaccgacg accgggtcat cgtggccgga tcttgcggcc cctcggcttg aacgaattgt
     2341 tagacattat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga caaattcttc
     2401 caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag cctgtctagc
     2461 ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt cggcagcgac
     2521 atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca acgtaagcac
     2581 tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta aggtttcatt
     2641 tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg ccgctggacc
     2701 taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat caatgtcgat
     2761 cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc caaattgcag
     2821 ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc
     2881 tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat
     2941 caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca aatcaatatc
     3001 actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg ccagcaacgt
     3061 cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg atacttcggc
     3121 gatcaccgct tccctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3181 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa tagctagctc
     3241 actcggtcgc tacgctccgg gcgtgagact gcggcgggcg ctgcggacac atacaaagtt
     3301 acccacagat tccgtggata agcaggggac taacatgtga ggcaaaacag cagggccgcg
     3361 ccggtggcgt ttttccatag gctccgccct cctgccagag ttcacataaa cagacgcttt
     3421 tccggtgcat ctgtgggagc cgtgaggctc aaccatgaat ctgacagtac gggcgaaacc
     3481 cgacaggact taaagatccc caccgtttcc ggcgggtcg
//
