LOCUS       sequence-310342-        3353 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term His-SUMO 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 3353)
  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 3353)
  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: 23 pCOLI_A_N-SUMO_C_STREP
FEATURES             Location/Qualifiers
     source          1..3353
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     503..522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     756..773
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    788..927
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(853..872)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1013..1035
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1029..1220)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1457..1476
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1457..1475
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1513..1535
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1555..1572
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1573..1860
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG"
     CDS             1873..1896
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(2032..2037,2038..2061,2062..2097,2098..2121)
                     /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(2215..2233)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2229..2276
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2341..2432
                     /gene="bla"
                     /label=AmpR promoter
     terminator      3297..3336
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
ORIGIN
        1 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
       61 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      121 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
      181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      661 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      721 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      781 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
      841 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
      901 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
      961 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     1021 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     1081 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     1141 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     1201 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catggggtaa
     1261 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     1321 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     1381 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ggcaccatgc
     1441 atcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt tccctccggc
     1501 tccggaaata attttgttta actttaagaa ggagatatac atatgggcag cagccatcat
     1561 catcatcatc actcggactc agaagtcaat caagaagcta agccagaggt caagccagaa
     1621 gtcaagcctg agactcacat caatttaaag gtgtccgatg gatcttcaga gatcttcttc
     1681 aagatcaaaa agaccactcc tttaagaagg ctgatggaag cgttcgctaa aagacagggt
     1741 aaggaaatgg actccttaag attcttgtac gacggtatta gaattcaagc tgatcagacc
     1801 cctgaagatt tggacatgga ggataacgat attattgagg ctcacagaga acagattggt
     1861 agcagcggat ctctggaagt tctgttccag gggcccggat cccggtccga catatgagcg
     1921 cgcggaattc aaaggcctac gtcgacgagc tcactagtcg cggccgcttt cgaatctaga
     1981 gcctgcagtc tcgacaagct tggaagcgga tctggatctg gaagcagcgc cagcgcttgg
     2041 agtcatcccc aatttgagaa aggtggaggt tccggaggtg gatcgggagg ttcggcgtgg
     2101 agccacccgc agttcgaaaa ataataagtc gagaagtact agaggatgta caacgcgtgc
     2161 gcgctgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     2221 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     2281 aaggaggaac tatatcggga ttagcccgcc taatgagcgg gctttttttt aattccccta
     2341 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2401 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2461 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctcgtga
     2521 aagtaaaaga cgcagaggac caattggggg cacgagtggg atacatagaa ctggacttga
     2581 atagcggtaa aatccttgag agttttcgcc ctgaagagcg ttttccaatg atgagcactt
     2641 tcaaagttct gctatgtgga gcagtattat cccgtgtaga tgcggggcaa gagcaactcg
     2701 gacgacgaat acactattcg cagaatgact tggttgaata ctccccagtg acagaaaagc
     2761 accttacgga cggaatgacg gtaagagaat tatgtagtgc cgccataacg atgagtgata
     2821 acactgcggc gaacttactt ctgacaacca tcggtggacc gaaggaatta accgcttttt
     2881 tgcacaatat gggagaccat gtaactcgcc ttgaccgttg ggaaccagaa ctgaatgaag
     2941 ccataccaaa cgacgagcga gacaccacaa tgcctgcggc aatggcaaca acattacgca
     3001 aactattaac tggcgaacta cttactctgg cttcacggca acaattaata gactggcttg
     3061 aagcggataa agttgcagga ccactactgc gttcggcact tcctgctggc tggtttattg
     3121 ctgataaatc tggggcagga gagcgtggtt cacggggtat cattgccgca cttggaccag
     3181 atggtaagcc ttcccgtatc gtagttatct acacgacggg tagtcaggca actatggacg
     3241 aacgaaatag acagattgct gaaatagggg cttcactgat taagcattgg taaaccgata
     3301 caattaaagg ctccttttgg agcctttttt tttggacgga ccggtagaaa aga
//
