pCJH10_pETcc2--E2Aa vector (V016024)

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LOCUS       62056_6465        3454 bp DNA     circular SYN 12-DEC-2019
DEFINITION  Expression vector pCJH10_pETcc2::E2Aa, complete sequence.
ACCESSION   MK910757
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3454)
  AUTHORS   Hartley CJ, Williams CC, Scoble JA, Churches QI, North A, French NG,
            Nebl T, Coia G, Warden AC, Simpson G, Frazer AR, Nixon Jensen C, 
            Turner NJ, Scott C.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   2  (bases 1 to 3454)
  AUTHORS   Hartley CJ, Williams CC, Scoble JA, Churches QI, North A, French NG,
            Nebl T, Coia G, Warden AC, Simpson G, Frazer AR, Nixon Jensen C, 
            Turner NJ, Scott C.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 3454)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (10-MAY-2019) Biocatalysis "
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-MAY-2019) Biocatalysis "
FEATURES             Location/Qualifiers
     source          1..3454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2..937
                     /codon_start=1
                     /transl_table=11
                     /product="carboxylesterase E2"
                     /label=carboxylesterase E2
                     /note="from Alicyclobacillus acidocaldarius (PDB 1U4N)"
                     /protein_id="QGT40748.1"
                     /translation="MMPLDPVIQQVLDQLNRMPAPDYKHLSAQQFRSQQSLFPPVKKEP
                     VAEVREFDMDLPGRTLKVRMYRPEGVEPPYPALVYYHGGGWVVGDLETHDPVCRVLAKD
                     GRAVVFSVDYRLAPEHKFPAAVEDAYDALQWIAERAADFHLDPARIAVGGDSAGGNLAA
                     VTSILAKERGGPALAFQLLIYPSTGYDPAHPPASIEENAEGYLLTGGMMLWFRDQYLNS
                     LEELTHPWFSPVLYPDLSGLPPAYIATAQYDPLRDVGKLYAEALNKAGVKVEIENFEDL
                     IHGFAQFYSLSPGATKALVRIAEKLRDALA"
     terminator      959..1006
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(1372..1400)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        1513..1617
                     /label=AmpR promoter
     CDS             1618..2475
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2649..3237
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        3317..3335
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3367..3389
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3408..3425
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             3435..3452
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"