LOCUS       sequence-316926-        3065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term 6xHis 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 3065)
  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 3065)
  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: 20 pCOLI_A_N-His_C_STREP
FEATURES             Location/Qualifiers
     source          1..3065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      282..321
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      352..940
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     841..860
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1094..1111
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1126..1265
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1191..1210)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1351..1373
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1367..1558)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1795..1814
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1795..1813
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1851..1873
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1893..1910
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1923..1946
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(2082..2087,2088..2111,2112..2147,2148..2171)
                     /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(2265..2283)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2279..2326
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2391..2482
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aactacttac tctggcttca cggcaacaat taatagactg gcttgaagcg gataaagttg
       61 caggaccact actgcgttcg gcacttcctg ctggctggtt tattgctgat aaatctgggg
      121 caggagagcg tggttcacgg ggtatcattg ccgcacttgg accagatggt aagccttccc
      181 gtatcgtagt tatctacacg acgggtagtc aggcaactat ggacgaacga aatagacaga
      241 ttgctgaaat aggggcttca ctgattaagc attggtaaac cgatacaatt aaaggctcct
      301 tttggagcct ttttttttgg acggaccggt agaaaagatc aaaggatctt cttgagatcc
      361 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
      421 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
      481 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
      541 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
      601 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
      661 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
      721 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
      781 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
      841 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
      901 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
      961 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     1021 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     1081 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     1141 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     1201 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     1261 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     1321 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     1381 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     1441 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     1501 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     1561 atgcctccgt gtaaggggga tttctgttca tggggtaatg ataccgatga aacgagagag
     1621 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     1681 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     1741 gcgcttcgtt aatacagatg taggtgttgg caccatgcat cgatcccgcg aaattaatac
     1801 gactcactat agggagacca caacggtttc cctccggctc cggaaataat tttgtttaac
     1861 tttaagaagg agatatacat atgggcagca gccatcatca tcatcatcac agcagcggat
     1921 ctctggaagt tctgttccag gggcccggat cccggtccga catatgagcg cgcggaattc
     1981 aaaggcctac gtcgacgagc tcactagtcg cggccgcttt cgaatctaga gcctgcagtc
     2041 tcgacaagct tggaagcgga tctggatctg gaagcagcgc cagcgcttgg agtcatcccc
     2101 aatttgagaa aggtggaggt tccggaggtg gatcgggagg ttcggcgtgg agccacccgc
     2161 agttcgaaaa ataataagtc gagaagtact agaggatgta caacgcgtgc gcgctgatcc
     2221 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     2281 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     2341 tatatcggga ttagcccgcc taatgagcgg gctttttttt aattccccta tttgtttatt
     2401 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2461 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2521 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctcgtga aagtaaaaga
     2581 cgcagaggac caattggggg cacgagtggg atacatagaa ctggacttga atagcggtaa
     2641 aatccttgag agttttcgcc ctgaagagcg ttttccaatg atgagcactt tcaaagttct
     2701 gctatgtgga gcagtattat cccgtgtaga tgcggggcaa gagcaactcg gacgacgaat
     2761 acactattcg cagaatgact tggttgaata ctccccagtg acagaaaagc accttacgga
     2821 cggaatgacg gtaagagaat tatgtagtgc cgccataacg atgagtgata acactgcggc
     2881 gaacttactt ctgacaacca tcggtggacc gaaggaatta accgcttttt tgcacaatat
     2941 gggagaccat gtaactcgcc ttgaccgttg ggaaccagaa ctgaatgaag ccataccaaa
     3001 cgacgagcga gacaccacaa tgcctgcggc aatggcaaca acattacgca aactattaac
     3061 tggcg
//
