LOCUS       sequence-318782-        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      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    775..914
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(840..859)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1000..1022
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1016..1207)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1444..1463
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1444..1462
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1500..1522
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1542..1559
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1560..1847
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG"
     CDS             1860..1883
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(2019..2024,2025..2048,2049..2084,2085..2108)
                     /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(2202..2220)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2216..2263
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2328..2419
                     /gene="bla"
                     /label=AmpR promoter
     terminator      3284..3323
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
      841 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
      901 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
      961 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     1021 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     1081 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     1141 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     1201 tggtcactga tgcctccgtg taagggggat ttctgttcat ggggtaatga taccgatgaa
     1261 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     1321 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     1381 tcaatgccag cgcttcgtta atacagatgt aggtgttggc accatgcatc gatcccgcga
     1441 aattaatacg actcactata gggagaccac aacggtttcc ctccggctcc ggaaataatt
     1501 ttgtttaact ttaagaagga gatatacata tgggcagcag ccatcatcat catcatcact
     1561 cggactcaga agtcaatcaa gaagctaagc cagaggtcaa gccagaagtc aagcctgaga
     1621 ctcacatcaa tttaaaggtg tccgatggat cttcagagat cttcttcaag atcaaaaaga
     1681 ccactccttt aagaaggctg atggaagcgt tcgctaaaag acagggtaag gaaatggact
     1741 ccttaagatt cttgtacgac ggtattagaa ttcaagctga tcagacccct gaagatttgg
     1801 acatggagga taacgatatt attgaggctc acagagaaca gattggtagc agcggatctc
     1861 tggaagttct gttccagggg cccggatccc ggtccgacat atgagcgcgc ggaattcaaa
     1921 ggcctacgtc gacgagctca ctagtcgcgg ccgctttcga atctagagcc tgcagtctcg
     1981 acaagcttgg aagcggatct ggatctggaa gcagcgccag cgcttggagt catccccaat
     2041 ttgagaaagg tggaggttcc ggaggtggat cgggaggttc ggcgtggagc cacccgcagt
     2101 tcgaaaaata ataagtcgag aagtactaga ggatgtacaa cgcgtgcgcg ctgatccggc
     2161 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     2221 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     2281 atcgggatta gcccgcctaa tgagcgggct tttttttaat tcccctattt gtttattttt
     2341 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2401 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2461 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctcgtgaaag taaaagacgc
     2521 agaggaccaa ttgggggcac gagtgggata catagaactg gacttgaata gcggtaaaat
     2581 ccttgagagt tttcgccctg aagagcgttt tccaatgatg agcactttca aagttctgct
     2641 atgtggagca gtattatccc gtgtagatgc ggggcaagag caactcggac gacgaataca
     2701 ctattcgcag aatgacttgg ttgaatactc cccagtgaca gaaaagcacc ttacggacgg
     2761 aatgacggta agagaattat gtagtgccgc cataacgatg agtgataaca ctgcggcgaa
     2821 cttacttctg acaaccatcg gtggaccgaa ggaattaacc gcttttttgc acaatatggg
     2881 agaccatgta actcgccttg accgttggga accagaactg aatgaagcca taccaaacga
     2941 cgagcgagac accacaatgc ctgcggcaat ggcaacaaca ttacgcaaac tattaactgg
     3001 cgaactactt actctggctt cacggcaaca attaatagac tggcttgaag cggataaagt
     3061 tgcaggacca ctactgcgtt cggcacttcc tgctggctgg tttattgctg ataaatctgg
     3121 ggcaggagag cgtggttcac ggggtatcat tgccgcactt ggaccagatg gtaagccttc
     3181 ccgtatcgta gttatctaca cgacgggtag tcaggcaact atggacgaac gaaatagaca
     3241 gattgctgaa ataggggctt cactgattaa gcattggtaa accgatacaa ttaaaggctc
     3301 cttttggagc cttttttttt ggacggaccg gtagaaaaga tcaaaggatc ttc
//
