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"
ORIGIN
        1 tatgatgccg ctggacccgg ttattcagca ggttctggat cagctgaacc gtatgccagc
       61 accggattat aaacatctga gcgcacagca gtttcgtagc cagcagagcc tgtttcctcc
      121 ggttaaaaaa gagccggttg cagaagttcg cgaatttgat atggatctgc caggtcgtac
      181 cctgaaagtt cgtatgtatc gtccggaagg tgttgaaccg ccttatccgg cactggttta
      241 ttatcatggt ggcggttggg tggttggtga tctggaaacc catgatcctg tttgtcgtgt
      301 tctggcaaaa gatggtcgtg cagttgtttt tagcgttgat tatcgcctgg caccggaaca
      361 taaatttccg gcagccgttg aagatgcgta tgatgcgctg cagtggattg ccgaacgtgc
      421 agcagatttt catctggacc ctgcccgtat tgccgttggt ggtgatagtg cgggtggtaa
      481 tctggcagcg gttaccagca ttctggccaa agaacgtggt ggtcctgcac tggcatttca
      541 gctgctgatc tatccgagca ccggttatga tccggcacat ccgccagcca gcattgaaga
      601 aaatgcagaa ggctacctgc tgacaggtgg tatgatgctg tggtttcgtg atcagtatct
      661 gaatagcctg gaagaactga cccatccgtg gtttagtccg gttctgtatc cggatctgag
      721 cggtctgcct ccggcatata ttgccaccgc acagtatgat cctctgcgtg acgttggtaa
      781 actgtatgca gaagccctga ataaagccgg tgttaaagtg gaaatcgaaa actttgaaga
      841 tctgatccat ggctttgccc agttttatag cctgagtccg ggtgcaacca aagcactggt
      901 tcgtattgca gaaaaactgc gggatgcact ggcctaataa ggatccgctg agcaataact
      961 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     1021 tatatccgga tatccacagg acgggtgtgg tcgccatgat cgcgtagtcg atagtggctc
     1081 caagtagcga agcgagcagg actgggcggc ggccaaagcg gtcggacagt gctccgagaa
     1141 cgggtgcgca tagaaattgc atcaacgcat atagcgctag cagcacgcca tagtgactgg
     1201 cgatgctgtc ggaatggacg atatcccgca agaggcccgg cagtaccggc ataaccaagc
     1261 ctatgcctac agcatccagg gtgacggtgc cgaggatgac gatgagcgca ttgttagatt
     1321 tcatacacgg tgcctgactg cgttagcaat ttaactgtga taaactaccg cattaaagct
     1381 tatcgatgat aagctgtcaa acatgagaat tcttgaagac gaaagggcct cgtgatacgc
     1441 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     1501 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1561 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     1621 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     1681 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     1741 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     1801 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1861 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1921 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     1981 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     2041 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2101 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2161 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     2221 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2281 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     2341 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2401 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2461 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     2521 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     2581 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2641 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2701 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2761 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     2821 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2881 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2941 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3001 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3061 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3121 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3181 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3241 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatggat
     3301 ctcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt tccctctaga
     3361 aataattttg tttaacttta agaaggagat ataccatggg cagcagccat catcatcatc
     3421 atcacagcag cggcctggtg ccgcgcggca gcca
//
