LOCUS       sequence-318633-        2578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term 2xStrepII 3C cleavable Kan resistant bacterial expression 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2578)
  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 2578)
  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: 37 pCOLI_K_STREP
FEATURES             Location/Qualifiers
     source          1..2578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     164..183
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        164..182
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             220..242
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(265..270,271..294,295..330,331..354)
                     /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"
     CDS             367..390
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(583..601)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      597..644
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(874..1689)
                     /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(874..1689)
                     /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     1597..1616
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1690..1781)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1797..2546)
                     /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(1807..1824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2047..2066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgtagcacc tgaagtcagc cccatacgat ataagttgta attctcatgt tagtcatgcc
       61 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
      121 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctaatacg actcactata
      181 gggagaccac aacggtttcc ctccggctcc ggaaataatt ttgtttaact ttaagaagga
      241 gatatacata tgggcagcag cgccagcgct tggagccacc cgcagttcga gaaaggtgga
      301 ggttccggag gtggatcggg aggttcggcg tggagccacc cgcagttcga aaaaagcagc
      361 ggatctctgg aagttctgtt ccaggggccc ggatcccggt ccgacatatg agcgcgcgga
      421 attcaaaggc ctacgtcgac gagctcacta gtcgcggccg ctttcgaatc tagagcctgc
      481 agtctcgaca agcttgtcga gaagtactag aggatgtaca acgcgtgcgc gctgatccgg
      541 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
      601 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa cctcaggcat
      661 ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac cagcaataga
      721 cataagcggc tatttaacga ccctgccctg aaccgacgac aagctgacga ccgggtctcc
      781 gcaagtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
      841 cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg
      901 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
      961 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     1021 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     1081 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     1141 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     1201 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcggtc
     1261 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     1321 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     1381 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     1441 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     1501 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     1561 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     1621 taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat
     1681 atggctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1741 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggcatgc agcgctcttc
     1801 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggtgtcagc
     1861 tcactcaaaa gcggtaatac ggttatccac agaatcaggg gataaagccg gaaagaacat
     1921 gtgagcaaaa agcaaagcac cggaagaagc caacgccgca ggcgtttttc cataggctcc
     1981 gcccccctga cgagcatcac aaaaatcgac gctcaagcca gaggtggcga aacccgacag
     2041 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2101 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2161 atagctcacg ctgttggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2221 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2281 ccaacccggt aagacacgac ttatcgccac tggcagcagc cattggtaac tgatttagag
     2341 gactttgtct tgaagttatg cacctgttaa ggctaaactg aaagaacaga ttttggtgag
     2401 tgcggtcctc caacccactt accttggttc aaagagttgg tagctcagcg aaccttgaga
     2461 aaaccaccgt tggtagcggt ggtttttctt tatttatgag atgatgaatc aatcggtcta
     2521 tcaagtcaac gaacagctat tccgttactc tagatttcag tgcaatttat ctcttcaa
//
