LOCUS       sequence-318781-        3683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term GST 3C cleavable, C--term 2xStrepII Amp resistant bacterial 
            expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3683)
  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 3683)
  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: 21 pCOLI_A_N-GST_C_STREP
FEATURES             Location/Qualifiers
     source          1..3683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     183..200
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    215..354
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(280..299)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     440..462
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(456..647)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     884..903
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        884..902
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             940..962
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             970..1623
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1581..1603
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1630..1653
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(1789..1794,1795..1818,1819..1854,1855..1878)
                     /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(1972..1990)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1986..2033
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2098..2189
                     /gene="bla"
                     /label=AmpR promoter
     terminator      3054..3093
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      join(3124..3683,1..29)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3613..3632
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
       61 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
      121 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
      181 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
      241 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
      301 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
      361 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
      421 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
      481 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
      541 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
      601 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
      661 taagggggat ttctgttcat ggggtaatga taccgatgaa acgagagagg atgctcacga
      721 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
      781 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
      841 atacagatgt aggtgttggc accatgcatc gatcccgcga aattaatacg actcactata
      901 gggagaccac aacggtttcc ctccggctcc ggaaataatt ttgtttaact ttaagaagga
      961 gatatacata tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact
     1021 cgacttcttt tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa
     1081 ggtgataaat ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat
     1141 tatattgatg gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac
     1201 aagcacaaca tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga
     1261 gcggttttgg atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact
     1321 ctcaaagttg attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta
     1381 tgtcataaaa catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac
     1441 gctcttgatg ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt
     1501 tgttttaaaa aacgtattga agctatccca caaattgata agtacttgaa atccagcaag
     1561 tatatagcat ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca
     1621 aaatcggatc tggaagttct gttccagggg cccggatccc ggtccgacat atgagcgcgc
     1681 ggaattcaaa ggcctacgtc gacgagctca ctagtcgcgg ccgctttcga atctagagcc
     1741 tgcagtctcg acaagcttgg aagcggatct ggatctggaa gcagcgccag cgcttggagt
     1801 catccccaat ttgagaaagg tggaggttcc ggaggtggat cgggaggttc ggcgtggagc
     1861 cacccgcagt tcgaaaaata ataagtcgag aagtactaga ggatgtacaa cgcgtgcgcg
     1921 ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     1981 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     2041 gaggaactat atcgggatta gcccgcctaa tgagcgggct tttttttaat tcccctattt
     2101 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2161 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2221 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctcgtgaaag
     2281 taaaagacgc agaggaccaa ttgggggcac gagtgggata catagaactg gacttgaata
     2341 gcggtaaaat ccttgagagt tttcgccctg aagagcgttt tccaatgatg agcactttca
     2401 aagttctgct atgtggagca gtattatccc gtgtagatgc ggggcaagag caactcggac
     2461 gacgaataca ctattcgcag aatgacttgg ttgaatactc cccagtgaca gaaaagcacc
     2521 ttacggacgg aatgacggta agagaattat gtagtgccgc cataacgatg agtgataaca
     2581 ctgcggcgaa cttacttctg acaaccatcg gtggaccgaa ggaattaacc gcttttttgc
     2641 acaatatggg agaccatgta actcgccttg accgttggga accagaactg aatgaagcca
     2701 taccaaacga cgagcgagac accacaatgc ctgcggcaat ggcaacaaca ttacgcaaac
     2761 tattaactgg cgaactactt actctggctt cacggcaaca attaatagac tggcttgaag
     2821 cggataaagt tgcaggacca ctactgcgtt cggcacttcc tgctggctgg tttattgctg
     2881 ataaatctgg ggcaggagag cgtggttcac ggggtatcat tgccgcactt ggaccagatg
     2941 gtaagccttc ccgtatcgta gttatctaca cgacgggtag tcaggcaact atggacgaac
     3001 gaaatagaca gattgctgaa ataggggctt cactgattaa gcattggtaa accgatacaa
     3061 ttaaaggctc cttttggagc cttttttttt ggacggaccg gtagaaaaga tcaaaggatc
     3121 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3181 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3241 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     3301 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3361 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3421 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3481 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3541 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3601 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3661 acttgagcgt cgatttttgt gat
//
