LOCUS       sequence-320903-        2522 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TEV protease plasmid for T7-driven E. coli cell-free protein 
            synthesis .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2522)
  AUTHORS   Dudley QM, Cai YM, Kallam K, Debreyne H, Carrasco Lopez JA, Patron 
            NJ
  TITLE     Biofoundry-assisted expression and characterization of plant 
            proteins.
  JOURNAL   Synth Biol (Oxf). 2021 Sep 11;6(1):ysab029. doi: 
            10.1093/synbio/ysab029. eCollection 2021.
  PUBMED    34693026
REFERENCE   2  (bases 1 to 2522)
  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: "Synth Biol 
            (Oxf)."; date: "2021-09-11"; volume: "6(1):ysab029. doi"; pages: " 
            10.1093/synbio/ysab029. eCollection 2021"
COMMENT     Sequence Label: pEPQDKN0729
FEATURES             Location/Qualifiers
     source          1..2522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      38..626
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     527..546
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     654..673
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        654..672
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             704..726
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             752..1459
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFI
                     VGIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPE
                     EPFQPVKEATQLMN"
     primer_bind     complement(1544..1562)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1558..1605
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1623..2438)
                     /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="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2346..2365
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2430..2449
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gagattttga gacacaacgt gagatcaaag gatcttcttg agatcctttt tttctgcgcg
       61 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
      121 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
      181 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
      241 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
      301 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
      361 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
      421 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
      481 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
      541 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
      601 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc gatcccgcga aattaatacg
      661 actcactata gggacaccac aacggtttcc ctctagaaat aattttgttt aactttaaga
      721 aggagatata catatggaga aaaaaataat gggtgagtct ctgttcaaag gcccacgcga
      781 ctataatccc atctcctcga cgatctgcca cttaaccaac gagtcggacg gacatacgac
      841 cagtctgtac ggcatcgggt tcggcccgtt tattatcact aataaacatc tgttccgtcg
      901 caacaacggt accttacttg tacagtctct tcacggcgtg tttaaagtga agaatactac
      961 tacccttcag cagcatctga tcgacggtcg cgatatgatt atcatccgta tgccgaaaga
     1021 ctttccgccc ttcccgcaga aattaaagtt ccgcgaaccc cagcgcgagg aacgtatttg
     1081 cttggttacc acaaattttc agacaaaatc aatgagctca atggttagcg atacgagctg
     1141 tacttttccg agcagtgacg gtatcttttg gaaacactgg atccagacga aagacggcca
     1201 atgcggatca ccgctggtga gtacccgtga cggctttatc gtgggcattc attccgccag
     1261 caactttact aataccaata actactttac ctcagtaccc aagaatttta tggagctgtt
     1321 aaccaaccaa gaagcacaac aatgggtgag cgggtggcgt ctgaacgccg atagcgttct
     1381 gtggggtggt cacaaggtgt ttatggtcaa gcctgaggaa ccgttccaac ctgtgaaaga
     1441 ggccacccag ttgatgaacg gttcgtaagt cgaccggctg ctaacaaagc ccgagcttcg
     1501 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     1561 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa agccaattct
     1621 gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     1681 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     1741 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     1801 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     1861 actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc
     1921 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     1981 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     2041 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     2101 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     2161 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     2221 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     2281 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     2341 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     2401 ggcttcgagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     2461 atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat
     2521 ca
//
