LOCUS       sequence-318775-        3308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term 2xStrepII 3C cleavable, C-term FKBP Amp resistant bacterial 
            expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3308)
  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 3308)
  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: 24 pCOLI_A_N_STREP_C_FKBP
FEATURES             Location/Qualifiers
     source          1..3308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..92
                     /gene="bla"
                     /label=AmpR promoter
     terminator      957..996
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1027..1615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1516..1535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1769..1786
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1801..1940
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1866..1885)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2026..2048
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2042..2233)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2470..2489
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2470..2488
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2526..2548
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(2571..2576,2577..2600,2601..2636,2637..2660)
                     /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             2673..2696
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3183..3201)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3197..3244
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
       61 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
      121 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctcgtga
      181 aagtaaaaga cgcagaggac caattggggg cacgagtggg atacatagaa ctggacttga
      241 atagcggtaa aatccttgag agttttcgcc ctgaagagcg ttttccaatg atgagcactt
      301 tcaaagttct gctatgtgga gcagtattat cccgtgtaga tgcggggcaa gagcaactcg
      361 gacgacgaat acactattcg cagaatgact tggttgaata ctccccagtg acagaaaagc
      421 accttacgga cggaatgacg gtaagagaat tatgtagtgc cgccataacg atgagtgata
      481 acactgcggc gaacttactt ctgacaacca tcggtggacc gaaggaatta accgcttttt
      541 tgcacaatat gggagaccat gtaactcgcc ttgaccgttg ggaaccagaa ctgaatgaag
      601 ccataccaaa cgacgagcga gacaccacaa tgcctgcggc aatggcaaca acattacgca
      661 aactattaac tggcgaacta cttactctgg cttcacggca acaattaata gactggcttg
      721 aagcggataa agttgcagga ccactactgc gttcggcact tcctgctggc tggtttattg
      781 ctgataaatc tggggcagga gagcgtggtt cacggggtat cattgccgca cttggaccag
      841 atggtaagcc ttcccgtatc gtagttatct acacgacggg tagtcaggca actatggacg
      901 aacgaaatag acagattgct gaaatagggg cttcactgat taagcattgg taaaccgata
      961 caattaaagg ctccttttgg agcctttttt tttggacgga ccggtagaaa agatcaaagg
     1021 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     1081 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     1141 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     1201 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     1261 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     1321 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     1381 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     1441 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     1501 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     1561 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     1621 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     1681 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     1741 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     1801 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     1861 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     1921 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     1981 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     2041 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     2101 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     2161 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     2221 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg taatgatacc
     2281 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
     2341 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
     2401 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttggcacca tgcatcgatc
     2461 ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctcc ggctccggaa
     2521 ataattttgt ttaactttaa gaaggagata tacatatggg cagcagcgcc agcgcttgga
     2581 gccacccgca gttcgagaaa ggtggaggtt ccggaggtgg atcgggaggt tcggcgtgga
     2641 gccacccgca gttcgaaaaa agcagcggat ctctggaagt tctgttccag gggcccggat
     2701 ccgacactag agcctccagt ctcgacaagc ttggaagcgg atctggatct ggaagcagcg
     2761 ccagcgcttc aggagttcag gtcgagacca tttcgcctgg agatggccgt acgttcccaa
     2821 agcgtggtca gacttgtgta gtgcattaca cgggaatgct tgaggacggt aaaaaggtgg
     2881 attcctcgcg cgaccgcaat aaaccgttta agttcatgct tggcaagcaa gaggtgattc
     2941 gtggctggga agagggtgtg gcccaaatgt ccgtcgggca acgcgcaaag ctgacgatca
     3001 gtccagacta tgcttacggc gcgactgggc atccagggat cattccgcca catgcaaccc
     3061 ttgtttttga cgttgagctg ctgaagttag aacttgtcga gaagtactag aggatgtaca
     3121 acgcgtgcgc gctgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     3181 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     3241 tttgctgaaa ggaggaacta tatcgggatt agcccgccta atgagcgggc ttttttttaa
     3301 ttccccta
//
