LOCUS       sequence-316929-        2503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term 6xHis 3C cleavable Kan resistant bacterial expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2503)
  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 2503)
  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: 34 pCOLI_K_His
FEATURES             Location/Qualifiers
     source          1..2503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     53..72
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        53..71
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             109..131
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             151..168
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             181..204
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(397..415)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      411..458
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(688..1503)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(688..1503)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1411..1430
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1504..1595)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1611..2360)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(1621..1638)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1861..1880)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctaatacga
       61 ctcactatag ggagaccaca acggtttccc tccggctccg gaaataattt tgtttaactt
      121 taagaaggag atatacatat gggcagcagc catcatcatc atcatcacag cagcggatct
      181 ctggaagttc tgttccaggg gcccggatcc cggtccgaca tatgagcgcg cggaattcaa
      241 aggcctacgt cgacgagctc actagtcgcg gccgctttcg aatctagagc ctgcagtctc
      301 gacaagcttg tcgagaagta ctagaggatg tacaacgcgt gcgcgctgat ccggctgcta
      361 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
      421 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaacctcag gcatttgaga
      481 agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa tagacataag
      541 cggctattta acgaccctgc cctgaaccga cgacaagctg acgaccgggt ctccgcaagt
      601 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
      661 aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg
      721 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
      781 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
      841 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
      901 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat
      961 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     1021 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc ggtcgctgtt
     1081 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     1141 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     1201 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     1261 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     1321 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     1381 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     1441 agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct
     1501 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1561 atacatattt gaatgtattt agaaaaataa acaaataggc atgcagcgct cttccgcttc
     1621 ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggtgt cagctcactc
     1681 aaaagcggta atacggttat ccacagaatc aggggataaa gccggaaaga acatgtgagc
     1741 aaaaagcaaa gcaccggaag aagccaacgc cgcaggcgtt tttccatagg ctccgccccc
     1801 ctgacgagca tcacaaaaat cgacgctcaa gccagaggtg gcgaaacccg acaggactat
     1861 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1921 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1981 cacgctgttg gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2041 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2101 cggtaagaca cgacttatcg ccactggcag cagccattgg taactgattt agaggacttt
     2161 gtcttgaagt tatgcacctg ttaaggctaa actgaaagaa cagattttgg tgagtgcggt
     2221 cctccaaccc acttaccttg gttcaaagag ttggtagctc agcgaacctt gagaaaacca
     2281 ccgttggtag cggtggtttt tctttattta tgagatgatg aatcaatcgg tctatcaagt
     2341 caacgaacag ctattccgtt actctagatt tcagtgcaat ttatctcttc aaatgtagca
     2401 cctgaagtca gccccatacg atataagttg taattctcat gttagtcatg ccccgcgccc
     2461 accggaagga gctgactggg ttgaaggctc tcaagggcat cgg
//
