LOCUS       sequence-310326-        2441 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 2441)
  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 2441)
  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..2441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(87..825)
                     /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     1035..1054
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1035..1053
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1091..1113
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(1182..1187,1188..1211,1212..1247,1248..1271)
                     /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(1365..1383)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1379..1426
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1597..2388)
                     /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(2389..2441,1..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 agcagttccc
      721 caactgactt aaccttcgat caaaccacct ccccaggtgg ttttttcgtt tacagggcaa
      781 aagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tactgaaccg
      841 ctctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga
      901 tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag ctgactgggt
      961 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     1021 taattgcgtt gcgctaatac gactcactat agggagacca caacggtttc cctccggctc
     1081 cggaaataat tttgtttaac tttaagaagg agactggatc cgaatctaga gcctgcagtc
     1141 tcgacaagct tggaagcgga tctggatctg gaagcagcgc cagcgcttgg agtcatcccc
     1201 aatttgagaa aggtggaggt tccggaggtg gatcgggagg ttcggcgtgg agccacccgc
     1261 agttcgaaaa ataataagtc gagaagtact agaggatgta caacgcgtgc gcgctgatcc
     1321 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     1381 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aacctcaggc
     1441 atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata
     1501 gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcat cgtggccgga
     1561 tcttgcggcc cctcggcttg aacgaattgt tagacattat ttgccgacta ccttggtgat
     1621 ctcgcctttc acgtagtgga caaattcttc caactgatct gcgcgcgagg ccaagcgatc
     1681 ttcttcttgt ccaagataag cctgtctagc ttcaagtatg acgggctgat actgggccgg
     1741 caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc cggttactgc
     1801 gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatcgccag cccagtcggg
     1861 cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct gttcaggaac
     1921 cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt ctcttgcttt
     1981 tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac ctgcaagaat
     2041 gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac gccacggaat
     2101 gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc tctctccagg
     2161 ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt catcaagcct
     2221 tacggtcacc gtaaccagca aatcaatatc actgtgtggc ttcaggccgc catccactgc
     2281 ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga tgacgccaac
     2341 tacctctgat agttgagtcg atacttcggc gatcaccgct tccctcatac tcttcctttt
     2401 tcaatattat tgaagcattt atcagggtta ttgtctcatg a
//
