LOCUS       sequence-318778-        4002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term MBP 3C cleavable Amp resistant bacterial expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4002)
  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 4002)
  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: 11 pCOLI_A_MBP
FEATURES             Location/Qualifiers
     source          1..4002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     167..186
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        167..185
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             223..245
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             259..276
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             286..1386
                     /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     1360..1383
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1396..1419
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1574..1592)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1588..1635
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1700..1791
                     /gene="bla"
                     /label=AmpR promoter
     terminator      2656..2695
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      2726..3314
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3215..3234
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3468..3485
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3500..3639
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3565..3584)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3725..3747
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3741..3932)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
       61 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
      121 ttaatacaga tgtaggtgtt ggcaccatgc atcgatcccg cgaaattaat acgactcact
      181 atagggagac cacaacggtt tccctccggc tccggaaata attttgttta actttaagaa
      241 ggagatatac atatggcaca ccatcaccac catcacagca gcggcatgaa aatcgaagaa
      301 ggtaaactgg taatctggat taacggcgat aaaggctata acggtctcgc tgaagtcggt
      361 aagaaattcg agaaagatac cggaattaaa gtcaccgttg agcatccgga taaactggaa
      421 gagaaattcc cacaggttgc ggcaactggc gatggccctg acattatctt ctgggcacac
      481 gaccgctttg gtggctacgc tcaatctggc ctgttggctg aaatcacccc ggacaaagcg
      541 ttccaggaca agctgtatcc gtttacctgg gatgccgtac gttacaacgg caagctgatt
      601 gcttacccga tcgctgttga agcgttatcg ctgatttata acaaagatct gctgccgaac
      661 ccgccaaaaa cctgggaaga gatcccggcg ctggataaag aactgaaagc gaaaggtaag
      721 agcgcgctga tgttcaacct gcaagaaccg tacttcacct ggccgctgat tgctgctgac
      781 gggggttatg cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat
      841 aacgctggcg cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa caaacacatg
      901 aatgcagaca ccgattactc catcgcagaa gctgccttta ataaaggcga aacagcgatg
      961 accatcaacg gcccgtgggc atggtccaac atcgacacca gcaaagtgaa ttatggtgta
     1021 acggtactgc cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca
     1081 ggtattaacg ccgccagtcc gaacaaagag ctggcaaaag agttcctcga aaactatctg
     1141 ctgactgatg aaggtctgga agcggttaat aaagacaaac cgctgggtgc cgtagcgctg
     1201 aagtcttacg aggaagagtt ggcgaaagat ccacgtattg ccgccactat ggaaaacgcc
     1261 cagaaaggtg aaatcatgcc gaacatcccg cagatgtccg ctttctggta tgccgtgcgt
     1321 actgcggtga tcaacgccgc cagcggtcgt cagactgtcg atgaagccct gaaagacgcg
     1381 cagactagca gcggtctgga agttctgttc caggggcccg gatccgtcga cgagctcact
     1441 agtcgcggcc gctttcgaat ctagagcctg cagtctcgac aagcttgtcg agaagtacta
     1501 gaggatgtac aacgcgtgcg cgctgatccg gctgctaaca aagcccgaaa ggaagctgag
     1561 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     1621 ttgaggggtt ttttgctgaa aggaggaact atatcgggat tagcccgcct aatgagcggg
     1681 ctttttttta attcccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1741 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     1801 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     1861 cacccagaaa cgctcgtgaa agtaaaagac gcagaggacc aattgggggc acgagtggga
     1921 tacatagaac tggacttgaa tagcggtaaa atccttgaga gttttcgccc tgaagagcgt
     1981 tttccaatga tgagcacttt caaagttctg ctatgtggag cagtattatc ccgtgtagat
     2041 gcggggcaag agcaactcgg acgacgaata cactattcgc agaatgactt ggttgaatac
     2101 tccccagtga cagaaaagca ccttacggac ggaatgacgg taagagaatt atgtagtgcc
     2161 gccataacga tgagtgataa cactgcggcg aacttacttc tgacaaccat cggtggaccg
     2221 aaggaattaa ccgctttttt gcacaatatg ggagaccatg taactcgcct tgaccgttgg
     2281 gaaccagaac tgaatgaagc cataccaaac gacgagcgag acaccacaat gcctgcggca
     2341 atggcaacaa cattacgcaa actattaact ggcgaactac ttactctggc ttcacggcaa
     2401 caattaatag actggcttga agcggataaa gttgcaggac cactactgcg ttcggcactt
     2461 cctgctggct ggtttattgc tgataaatct ggggcaggag agcgtggttc acggggtatc
     2521 attgccgcac ttggaccaga tggtaagcct tcccgtatcg tagttatcta cacgacgggt
     2581 agtcaggcaa ctatggacga acgaaataga cagattgctg aaataggggc ttcactgatt
     2641 aagcattggt aaaccgatac aattaaaggc tccttttgga gccttttttt ttggacggac
     2701 cggtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2761 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2821 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2881 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2941 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3001 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3061 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3121 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3181 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3241 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3301 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3361 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3421 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3481 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3541 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3601 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     3661 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3721 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     3781 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     3841 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     3901 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     3961 ttcatggggt aatgataccg atgaaacgag agaggatgct ca
//
