LOCUS       sequence-316942-        2791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term 6xHis SUMO 3C cleavable Kan resistant bacterial expression 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2791)
  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 2791)
  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: 39 pCOLI_K_SUMO
FEATURES             Location/Qualifiers
     source          1..2791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(96..845)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(106..123)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(346..365)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1041..1060
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1041..1059
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1097..1119
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1139..1156
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1157..1444
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG"
     CDS             1457..1480
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1673..1691)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1687..1734
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1964..2779)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1964..2779)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2687..2706
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(join(2780..2791,1..80))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
       61 tatttagaaa aataaacaaa taggcatgca gcgctcttcc gcttcctcgc tcactgactc
      121 gctacgctcg gtcgttcgac tgcggcgagc ggtgtcagct cactcaaaag cggtaatacg
      181 gttatccaca gaatcagggg ataaagccgg aaagaacatg tgagcaaaaa gcaaagcacc
      241 ggaagaagcc aacgccgcag gcgtttttcc ataggctccg cccccctgac gagcatcaca
      301 aaaatcgacg ctcaagccag aggtggcgaa acccgacagg actataaaga taccaggcgt
      361 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      421 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgttggtatc
      481 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      541 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      601 tatcgccact ggcagcagcc attggtaact gatttagagg actttgtctt gaagttatgc
      661 acctgttaag gctaaactga aagaacagat tttggtgagt gcggtcctcc aacccactta
      721 ccttggttca aagagttggt agctcagcga accttgagaa aaccaccgtt ggtagcggtg
      781 gtttttcttt atttatgaga tgatgaatca atcggtctat caagtcaacg aacagctatt
      841 ccgttactct agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc
      901 atacgatata agttgtaatt ctcatgttag tcatgccccg cgcccaccgg aaggagctga
      961 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     1021 ttacattaat tgcgttgcgc taatacgact cactataggg agaccacaac ggtttccctc
     1081 cggctccgga aataattttg tttaacttta agaaggagat atacatatgg gcagcagcca
     1141 tcatcatcat catcactcgg actcagaagt caatcaagaa gctaagccag aggtcaagcc
     1201 agaagtcaag cctgagactc acatcaattt aaaggtgtcc gatggatctt cagagatctt
     1261 cttcaagatc aaaaagacca ctcctttaag aaggctgatg gaagcgttcg ctaaaagaca
     1321 gggtaaggaa atggactcct taagattctt gtacgacggt attagaattc aagctgatca
     1381 gacccctgaa gatttggaca tggaggataa cgatattatt gaggctcaca gagaacagat
     1441 tggtagcagc ggatctctgg aagttctgtt ccaggggccc ggatcccggt ccgacatatg
     1501 agcgcgcgga attcaaaggc ctacgtcgac gagctcacta gtcgcggccg ctttcgaatc
     1561 tagagcctgc agtctcgaca agcttgtcga gaagtactag aggatgtaca acgcgtgcgc
     1621 gctgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     1681 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     1741 cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
     1801 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac aagctgacga
     1861 ccgggtctcc gcaagtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1921 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
     1981 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     2041 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     2101 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     2161 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     2221 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     2281 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     2341 atacgcggtc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     2401 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     2461 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     2521 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     2581 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     2641 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     2701 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
     2761 cccgttgaat atggctcata ctcttccttt t
//
