LOCUS       sequence-316928-        4129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term MBP 3C cleavable, C--term 2xStrepII Amp resistant bacterial 
            expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4129)
  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 4129)
  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: 22 pCOLI_A_N-MBP_C_STREP
FEATURES             Location/Qualifiers
     source          1..4129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     41..60
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        41..59
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             97..119
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             133..150
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             160..1260
                     /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     1234..1257
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1270..1293
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(1392..1397,1398..1421,1422..1457,1458..1481)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     primer_bind     complement(1575..1593)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1589..1636
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1701..1792
                     /gene="bla"
                     /label=AmpR promoter
     terminator      2657..2696
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      2727..3315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3216..3235
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3469..3486
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3501..3640
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3566..3585)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3726..3748
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3742..3933)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cagatgtagg tgttggcacc atgcatcgat cccgcgaaat taatacgact cactataggg
       61 agaccacaac ggtttccctc cggctccgga aataattttg tttaacttta agaaggagat
      121 atacatatgg cacaccatca ccaccatcac agcagcggca tgaaaatcga agaaggtaaa
      181 ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt cggtaagaaa
      241 ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact ggaagagaaa
      301 ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc acacgaccgc
      361 tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa agcgttccag
      421 gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct gattgcttac
      481 ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc gaacccgcca
      541 aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg taagagcgcg
      601 ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc tgacgggggt
      661 tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt ggataacgct
      721 ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca catgaatgca
      781 gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc gatgaccatc
      841 aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg tgtaacggta
      901 ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt
      961 aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta tctgctgact
     1021 gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc gctgaagtct
     1081 tacgaggaag agttggcgaa agatccacgt attgccgcca ctatggaaaa cgcccagaaa
     1141 ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt gcgtactgcg
     1201 gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga cgcgcagact
     1261 agcagcggtc tggaagttct gttccagggg cccggatccg tcgacgagct cactagtcgc
     1321 ggccgctttc gaatctagag cctgcaggtc tcgacaagct tggaagcgga tctggatctg
     1381 gaagcagcgc cagcgcttgg agtcatcccc aatttgagaa aggtggaggt tccggaggtg
     1441 gatcgggagg ttcggcgtgg agccacccgc agttcgaaaa ataataagtc gagaagtact
     1501 agaggatgta caacgcgtgc gcgctgatcc ggctgctaac aaagcccgaa aggaagctga
     1561 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     1621 cttgaggggt tttttgctga aaggaggaac tatatcggga ttagcccgcc taatgagcgg
     1681 gctttttttt aattccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1741 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1801 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1861 tcacccagaa acgctcgtga aagtaaaaga cgcagaggac caattggggg cacgagtggg
     1921 atacatagaa ctggacttga atagcggtaa aatccttgag agttttcgcc ctgaagagcg
     1981 ttttccaatg atgagcactt tcaaagttct gctatgtgga gcagtattat cccgtgtaga
     2041 tgcggggcaa gagcaactcg gacgacgaat acactattcg cagaatgact tggttgaata
     2101 ctccccagtg acagaaaagc accttacgga cggaatgacg gtaagagaat tatgtagtgc
     2161 cgccataacg atgagtgata acactgcggc gaacttactt ctgacaacca tcggtggacc
     2221 gaaggaatta accgcttttt tgcacaatat gggagaccat gtaactcgcc ttgaccgttg
     2281 ggaaccagaa ctgaatgaag ccataccaaa cgacgagcga gacaccacaa tgcctgcggc
     2341 aatggcaaca acattacgca aactattaac tggcgaacta cttactctgg cttcacggca
     2401 acaattaata gactggcttg aagcggataa agttgcagga ccactactgc gttcggcact
     2461 tcctgctggc tggtttattg ctgataaatc tggggcagga gagcgtggtt cacggggtat
     2521 cattgccgca cttggaccag atggtaagcc ttcccgtatc gtagttatct acacgacggg
     2581 tagtcaggca actatggacg aacgaaatag acagattgct gaaatagggg cttcactgat
     2641 taagcattgg taaaccgata caattaaagg ctccttttgg agcctttttt tttggacgga
     2701 ccggtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2761 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2821 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     2881 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2941 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3001 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3061 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3121 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3181 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3241 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3301 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3361 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3421 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3481 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3541 tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     3601 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     3661 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3721 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     3781 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     3841 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     3901 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
     3961 gttcatgggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     4021 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     4081 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaata
//
