LOCUS       Exported                2328 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 nsp8.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046790
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2328)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2328)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2328
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    39..63
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             78..100
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             705..725
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             726..749
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      866..1454
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1355..1374
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1560..2219
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1626..1645)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2290..2324
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
       61 tcccctctag aaataatttt gtttaacttt aagaaggaga ctcttctatg gccattgcga
      121 gcgaattttc aagtcttcca tcctatgctg cattcgctac tgcacaagag gcttacgaac
      181 aggcggtggc caacggcgac tcggaagtcg ttctgaagaa actgaaaaaa tctttaaatg
      241 ttgcgaaaag cgaatttgat cgtgacgctg cgatgcagcg caagttggag aaaatggctg
      301 atcaggcaat gacgcaaatg tacaaacagg ctcgttccga agacaaaagg gccaaagtaa
      361 cctctgctat gcagaccatg ctgttcacaa tgctgcgcaa gctggataac gacgcgctca
      421 acaacatcat caacaatgca cgtgatggtt gcgtacctct caacattatc ccgcttacta
      481 ctgctgcaaa actgatggtg gttataccgg actacaacac ctataagaac acctgtgatg
      541 ggacgacttt cacctacgcc agcgcactgt gggaaatcca gcaggttgtc gatgctgaca
      601 gtaaaatcgt tcagctgagc gagatttcta tggacaattc tccgaacctg gcgtggccac
      661 tgatcgtaac tgctctgcgt gcgaatagcg cggtcaaatt acaagagaac ctgtactttc
      721 aaagcgacta caaggatgat gatgacaaat aaccggctta tcggtcagtt tcacctgatt
      781 tacgtaaaaa cccgcttcgg cgggtttttg cttttggagg ggcagaaaga tgaatgactg
      841 tccacgacgc tatacccaaa agaaattgag atcctttttt tctgcgcgta atctgctgct
      901 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      961 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     1021 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1081 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1141 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1201 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     1261 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1321 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     1381 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     1441 ggagcctatg gaaattgatc gggcacgtaa gaggttccaa ctttcaccat aatgaaataa
     1501 gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa
     1561 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
     1621 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
     1681 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
     1741 acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg
     1801 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
     1861 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
     1921 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
     1981 atatgttttt cgtttcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
     2041 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg
     2101 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg
     2161 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat
     2221 ttgatatcga gctcgcttgg actcctgttg atagatccag taatgacctc agaactccat
     2281 ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggt
//
