LOCUS       sequence-319345-        2442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-term 2xStrepII uncleavable Amp resistant bacterial expression 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2442)
  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 2442)
  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: 29 pCOLI_S_C_STREP
FEATURES             Location/Qualifiers
     source          1..2442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(87..826)
                     /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     1036..1055
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1036..1054
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1092..1114
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(1183..1188,1189..1212,1213..1248,1249..1272)
                     /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(1366..1384)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1380..1427
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1598..2389)
                     /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(join(2390..2442,1..39))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcggatacat atttgaatgt atttagaaaa ataaacaaat agctagctca ctcggtcgct
       61 acgctccggg cgtgagactg cggcgggcgc tgcggacaca tacaaagtta cccacagatt
      121 ccgtggataa gcaggggact aacatgtgag gcaaaacagc agggccgcgc cggtggcgtt
      181 tttccatagg ctccgccctc ctgccagagt tcacataaac agacgctttt ccggtgcatc
      241 tgtgggagcc gtgaggctca accatgaatc tgacagtacg ggcgaaaccc gacaggactt
      301 aaagatcccc accgtttccg gcgggtcgct ccctcttgcg ctctcctgtt ccgaccctgc
      361 cgtttaccgg atacctgttc cgcctttctc ccttacggga agtgtggcgc tttctcatag
      421 ctcacacact ggtatctcgg ctcggtgtag gtcgttcgct ccaagctggg ctgtaagcaa
      481 gaactccccg ttcagcccga ctgctgcgcc ttatccggta actgttcact tgagtccaac
      541 ccggaaaagc acggtaaaac gccactggca gcagccattg gtaactggga gttcgcagag
      601 gatttgttta gctaaacacg cggttgctct tgaagtgtgc gccaaagtcc ggctacactg
      661 gaaggacaga tttggttgct gtgctctgcg aaagccagtt accacggtta agncagttss
      721 ssaactgnct taaccttcga tcaaaccacc tccccaggtg gttttttcgt ttacagggca
      781 aaagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctactgaacc
      841 gctctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg
      901 atataagttg taattctcat gttagtcatg ccccgcgccc accggaagga gctgactggg
      961 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     1021 ttaattgcgt tgcgctaata cgactcacta tagggagacc acaacggttt ccctccggct
     1081 ccggaaataa ttttgtttaa ctttaagaag gagactggat ccgaatctag agcctgcagt
     1141 ctcgacaagc ttggaagcgg atctggatct ggaagcagcg ccagcgcttg gagtcatccc
     1201 caatttgaga aaggtggagg ttccggaggt ggatcgggag gttcggcgtg gagccacccg
     1261 cagttcgaaa aataataagt cgagaagtac tagaggatgt acaacgcgtg cgcgctgatc
     1321 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     1381 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaacctcagg
     1441 catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat
     1501 agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtca tcgtggccgg
     1561 atcttgcggc ccctcggctt gaacgaattg ttagacatta tttgccgact accttggtga
     1621 tctcgccttt cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag gccaagcgat
     1681 cttcttcttg tccaagataa gcctgtctag cttcaagtat gacgggctga tactgggccg
     1741 gcaggcgctc cattgcccag tcggcagcga catccttcgg cgcgattttg ccggttactg
     1801 cgctgtacca aatgcgggac aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg
     1861 gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa
     1921 ccggatcaaa gagttcctcc gccgctggac ctaccaaggc aacgctatgt tctcttgctt
     1981 ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa
     2041 tgtcattgcg ctgccattct ccaaattgca gttcgcgctt agctggataa cgccacggaa
     2101 tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg gagaatctcg ctctctccag
     2161 gggaagccga agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc
     2221 ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg ccatccactg
     2281 cggagccgta caaatgtacg gccagcaacg tcggttcgag atggcgctcg atgacgccaa
     2341 ctacctctga tagttgagtc gatacttcgg cgatcaccgc ttccctcata ctcttccttt
     2401 ttcaatatta ttgaagcatt tatcagggtt attgtctcat ga
//
