LOCUS       sequence-316800-        3014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-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 3014)
  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 3014)
  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: 5 pCOLI_A_STREP
FEATURES             Location/Qualifiers
     source          1..3014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     342..361
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     595..612
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    627..766
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(692..711)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     852..874
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(868..1059)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1296..1315
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1296..1314
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1352..1374
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(1397..1402,1403..1426,1427..1462,1463..1486)
                     /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             1499..1522
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1715..1733)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1729..1776
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1841..1932
                     /gene="bla"
                     /label=AmpR promoter
     terminator      2797..2836
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      join(2867..3014,1..441)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
       61 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
      121 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
      181 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
      241 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
      301 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
      361 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
      421 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
      481 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
      541 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
      601 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
      661 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
      721 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
      781 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
      841 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
      901 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
      961 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     1021 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     1081 atttctgttc atggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     1141 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     1201 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     1261 gtaggtgttg gcaccatgca tcgatcccgc gaaattaata cgactcacta tagggagacc
     1321 acaacggttt ccctccggct ccggaaataa ttttgtttaa ctttaagaag gagatataca
     1381 tatgggcagc agcgccagcg cttggagcca cccgcagttc gagaaaggtg gaggttccgg
     1441 aggtggatcg ggaggttcgg cgtggagcca cccgcagttc gaaaaaagca gcggatctct
     1501 ggaagttctg ttccaggggc ccggatcccg gtccgacata tgagcgcgcg gaattcaaag
     1561 gcctacgtcg acgagctcac tagtcgcggc cgctttcgaa tctagagcct gcagtctcga
     1621 caagcttgtc gagaagtact agaggatgta caacgcgtgc gcgctgatcc ggctgctaac
     1681 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     1741 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatcggga
     1801 ttagcccgcc taatgagcgg gctttttttt aattccccta tttgtttatt tttctaaata
     1861 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1921 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     1981 ttttgccttc ctgtttttgc tcacccagaa acgctcgtga aagtaaaaga cgcagaggac
     2041 caattggggg cacgagtggg atacatagaa ctggacttga atagcggtaa aatccttgag
     2101 agttttcgcc ctgaagagcg ttttccaatg atgagcactt tcaaagttct gctatgtgga
     2161 gcagtattat cccgtgtaga tgcggggcaa gagcaactcg gacgacgaat acactattcg
     2221 cagaatgact tggttgaata ctccccagtg acagaaaagc accttacgga cggaatgacg
     2281 gtaagagaat tatgtagtgc cgccataacg atgagtgata acactgcggc gaacttactt
     2341 ctgacaacca tcggtggacc gaaggaatta accgcttttt tgcacaatat gggagaccat
     2401 gtaactcgcc ttgaccgttg ggaaccagaa ctgaatgaag ccataccaaa cgacgagcga
     2461 gacaccacaa tgcctgcggc aatggcaaca acattacgca aactattaac tggcgaacta
     2521 cttactctgg cttcacggca acaattaata gactggcttg aagcggataa agttgcagga
     2581 ccactactgc gttcggcact tcctgctggc tggtttattg ctgataaatc tggggcagga
     2641 gagcgtggtt cacggggtat cattgccgca cttggaccag atggtaagcc ttcccgtatc
     2701 gtagttatct acacgacggg tagtcaggca actatggacg aacgaaatag acagattgct
     2761 gaaatagggg cttcactgat taagcattgg taaaccgata caattaaagg ctccttttgg
     2821 agcctttttt tttggacgga ccggtagaaa agatcaaagg atcttcttga gatccttttt
     2881 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2941 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3001 taccaaatac tgtc
//
