LOCUS       sequence-318630-        2489 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-term 2xStrepII uncleavable Kan resistant bacterial expression 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2489)
  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 2489)
  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: 38 pCOLI_K_ C_STREP
FEATURES             Location/Qualifiers
     source          1..2489
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(180..227)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     223..241
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(join(335..358,359..394,395..418,419..424))
                     /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"
     RBS             493..515
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(552..571)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(553..571)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      767..1516
                     /direction=RIGHT
                     /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     1247..1266
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1489..1506
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1532..1623
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1624..2439
                     /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             1624..2439
                     /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     complement(1697..1716)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 aaataggggt tccgcgcaca tttccccgaa aagtgccact tgcggagacc cggtcgtcag
       61 cttgtcgtcg gttcagggca gggtcgttaa atagccgctt atgtctattg ctggtttacc
      121 ggtttattga ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg aggtttcagc
      181 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg
      241 gtggcagcag ccaactcagc ttcctttcgg gctttgttag cagccggatc agcgcgcacg
      301 cgttgtacat cctctagtac ttctcgactt attatttttc gaactgcggg tggctccacg
      361 ccgaacctcc cgatccacct ccggaacctc cacctttctc aaattgggga tgactccaag
      421 cgctggcgct gcttccagat ccagatccgc ttccaagctt gtcgagactg caggctctag
      481 attcggatcc agtctccttc ttaaagttaa acaaaattat ttccggagcc ggagggaaac
      541 cgttgtggtc tccctatagt gagtcgtatt agcgcaacgc aattaatgta agttagctca
      601 ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
      661 tccggtgggc gcggggcatg actaacatga gaattacaac ttatatcgta tggggctgac
      721 ttcaggtgct acatttgaag agataaattg cactgaaatc tagagtaacg gaatagctgt
      781 tcgttgactt gatagaccga ttgattcatc atctcataaa taaagaaaaa ccaccgctac
      841 caacggtggt tttctcaagg ttcgctgagc taccaactct ttgaaccaag gtaagtgggt
      901 tggaggaccg cactcaccaa aatctgttct ttcagtttag ccttaacagg tgcataactt
      961 caagacaaag tcctctaaat cagttaccaa tggctgctgc cagtggcgat aagtcgtgtc
     1021 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     1081 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agataccaac
     1141 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1201 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1261 atctttatag tcctgtcggg tttcgccacc tctggcttga gcgtcgattt ttgtgatgct
     1321 cgtcaggggg gcggagccta tggaaaaacg cctgcggcgt tggcttcttc cggtgctttg
     1381 ctttttgctc acatgttctt tccggcttta tcccctgatt ctgtggataa ccgtattacc
     1441 gcttttgagt gagctgacac cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg
     1501 agcgaggaag cggaagagcg ctgcatgcct atttgtttat ttttctaaat acattcaaat
     1561 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1621 agtatgagcc atattcaacg ggaaacgtct tgctctaggc cgcgattaaa ttccaacatg
     1681 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     1741 atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     1801 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     1861 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     1921 gcgatccccg ggaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     1981 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     2041 ccttttaaca gcgaccgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     2101 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     2161 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     2221 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     2281 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     2341 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2401 aaattgcagt ttcatttgat gctcgatgag tttttctaag aattaattca tgagcggata
     2461 catatttgaa tgtatttaga aaaataaac
//
