LOCUS       sequence-316890-        2955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-term 2xStrepII 3C cleavable Amp resistant bacterial expression 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2955)
  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 2955)
  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: 7 pCOLI_A_3C_C_STREP
FEATURES             Location/Qualifiers
     source          1..2955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     437..456
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     690..707
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    722..861
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(787..806)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     947..969
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(963..1154)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1391..1410
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1391..1409
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1447..1469
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1544..1567
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(1574..1597,1598..1633,1634..1657)
                     /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="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     primer_bind     complement(1751..1769)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1765..1812
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1877..1968
                     /gene="bla"
                     /label=AmpR promoter
     terminator      2833..2872
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      join(2903..2955,1..536)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
       61 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
      121 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      181 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      241 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      301 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      361 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      421 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
      481 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
      541 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
      601 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
      661 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
      721 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac
      781 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
      841 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
      901 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
      961 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     1021 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     1081 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     1141 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg gtaatgatac
     1201 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     1261 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     1321 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttggcacc atgcatcgat
     1381 cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc cggctccgga
     1441 aataattttg tttaacttta agaaggagac tggatccgaa tctagagcct gcagtctcga
     1501 caagcttgga agcggatctg gatctggaag cagcgccagc gctctggaag ttctgttcca
     1561 ggggcccggt agctggagtc atccccaatt tgagaaaggt ggaggttccg gaggtggatc
     1621 gggaggttcg gcgtggagcc acccgcagtt cgaaaaataa taagtcgaga agtactagag
     1681 gatgtacaac gcgtgcgcgc tgatccggct gctaacaaag cccgaaagga agctgagttg
     1741 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     1801 aggggttttt tgctgaaagg aggaactata tcgggattag cccgcctaat gagcgggctt
     1861 ttttttaatt cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     1921 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     1981 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2041 ccagaaacgc tcgtgaaagt aaaagacgca gaggaccaat tgggggcacg agtgggatac
     2101 atagaactgg acttgaatag cggtaaaatc cttgagagtt ttcgccctga agagcgtttt
     2161 ccaatgatga gcactttcaa agttctgcta tgtggagcag tattatcccg tgtagatgcg
     2221 gggcaagagc aactcggacg acgaatacac tattcgcaga atgacttggt tgaatactcc
     2281 ccagtgacag aaaagcacct tacggacgga atgatggtaa gagaattatg tagtgccgcc
     2341 ataacgatga gtgataacac tgcggcgaac ttacttctga caaccatcgg tggaccgaag
     2401 gaattaaccg cttttttgca caatatggga gaccatgtaa ctcgccttga ccgttgggaa
     2461 ccagaactga atgaagccat accaaacgac gagcgagaca ccacaatgcc tgcggcaatg
     2521 gcaacaacat tacgcaaact attaactggc gaactactta ctctggcttc acggcaacaa
     2581 ttaatagact ggcttgaagc ggataaagtt gcaggaccac tactgcgttc ggcacttcct
     2641 gctggctggt ttattgctga taaatctggg gcaggagagc gtggttcacg gggtatcatt
     2701 gccgcacttg gaccagatgg taagccttcc cgtatcgtag ttatctacac gacgggtagt
     2761 caggcaacta tggacgaacg aaatagacag attgctgaaa taggggcttc actgattaag
     2821 cattggtaaa ccgatacaat taaaggctcc ttttggagcc tttttttttg gacggaccgg
     2881 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2941 aaacaaaaaa accac
//
