LOCUS       sequence-319362-        2530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term 2xStrepII 3C cleavable Spec resistant bacterial expression 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2530)
  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 2530)
  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: 28 pCOLI_S_STREP
FEATURES             Location/Qualifiers
     source          1..2530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(39..777)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     987..1006
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        987..1005
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1043..1065
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(1088..1093,1094..1117,1118..1153,1154..1177)
                     /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             1190..1213
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1406..1424)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1420..1467
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1638..2429)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2430..2521)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cactcggtcg ctacgctccg ggcgtgagac tgcggcgggc gctgcggaca catacaaagt
       61 tacccacaga ttccgtggat aagcagggga ctaacatgtg aggcaaaaca gcagggccgc
      121 gccggtggcg tttttccata ggctccgccc tcctgccaga gttcacataa acagacgctt
      181 ttccggtgca tctgtgggag ccgtgaggct caaccatgaa tctgacagta cgggcgaaac
      241 ccgacaggac ttaaagatcc ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg
      301 ttccgaccct gccgtttacc ggatacctgt tccgcctttc tcccttacgg gaagtgtggc
      361 gctttctcat agctcacaca ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg
      421 ggctgtaagc aagaactccc cgttcagccc gactgctgcg ccttatccgg taactgttca
      481 cttgagtcca acccggaaaa gcacggtaaa acgccactgg cagcagccat tggtaactgg
      541 gagttcgcag aggatttgtt tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt
      601 ccggctacac tggaaggaca gatttggttg ctgtgctctg cgaaagccag ttaccacggt
      661 taagcagtts sssaactgnc ttaaccttcg atcaaaccac ctccccaggt ggttttttcg
      721 wwtacagggc aaaagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      781 tctactgaac cgctctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
      841 agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg
      901 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
      961 gctaacttac attaattgcg ttgcgctaat acgactcact atagggagac cacaacggtt
     1021 tccctccggc tccggaaata attttgttta actttaagaa ggagatatac atatgggcag
     1081 cagcgccagc gcttggagcc acccgcagtt cgagaaaggt ggaggttccg gaggtggatc
     1141 gggaggttcg gcgtggagcc acccgcagtt cgaaaaaagc agcggatctc tggaagttct
     1201 gttccagggg cccggatccc ggtccgacat atgagcgcgc ggaattcaaa ggcctacgtc
     1261 gacgagctca ctagtcgcgg ccgctttcga atctagagcc tgcagtctcg acaagcttgt
     1321 cgagaagtac tagaggatgt acaacgcgtg cgcgctgatc cggctgctaa caaagcccga
     1381 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     1441 tctaaacggg tcttgagggg ttttttgctg aaacctcagg catttgagaa gcacacggtc
     1501 acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa
     1561 cgaccctgcc ctgaaccgac gaccgggtca tcgtggccgg atcttgcggc ccctcggctt
     1621 gaacgaattg ttagacatta tttgccgact accttggtga tctcgccttt cacgtagtgg
     1681 acaaattctt ccaactgatc tgcgcgcgag gccaagcgat cttcttcttg tccaagataa
     1741 gcctgtctag cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag
     1801 tcggcagcga catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac
     1861 aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt
     1921 aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc
     1981 gccgctggac ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga
     2041 tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct
     2101 ccaaattgca gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca
     2161 atggtgactt ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa
     2221 aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc
     2281 aaatcaatat cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg
     2341 gccagcaacg tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc
     2401 gatacttcgg cgatcaccgc ttccctcata ctcttccttt ttcaatatta ttgaagcatt
     2461 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     2521 atagctagct
//
