LOCUS       Exported                2339 bp ds-DNA     circular SYN 01-APR-2022
DEFINITION  Bacterial expression plasmid for SARS-CoV-2 nsp8.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046834
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2339)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2339)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 1, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2339
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     RBS             1..23
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             628..648
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             649..675
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      796..1384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1285..1304
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1490..2149
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1556..1575)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2220..2254
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     primer_bind     2282..2301
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2282..2300
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2301..2325
                     /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)."
ORIGIN
        1 tttgtttaac tttaagaagg agactcttct atggccattg cgagcgaatt ttcaagtctt
       61 ccatcctatg ctgcattcgc tactgcacaa gaggcttacg aacaggcggt ggccaacggc
      121 gactcggaag tcgttctgaa gaaactgaaa aaatctttaa atgttgcgaa aagcgaattt
      181 gatcgtgacg ctgcgatgca gcgcaagttg gagaaaatgg ctgatcaggc aatgacgcaa
      241 atgtacaaac aggctcgttc cgaagacaaa agggccaaag taacctctgc tatgcagacc
      301 atgctgttca caatgctgcg caagctggat aacgacgcgc tcaacaacat catcaacaat
      361 gcacgtgatg gttgcgtacc tctcaacatt atcccgctta ctactgctgc aaaactgatg
      421 gtggttatac cggactacaa cacctataag aacacctgtg atgggacgac tttcacctac
      481 gccagcgcac tgtgggaaat ccagcaggtt gtcgatgctg acagtaaaat cgttcagctg
      541 agcgagattt ctatggacaa ttctccgaac ctggcgtggc cactgatcgt aactgctctg
      601 cgtgcgaata gcgcggtcaa attacaagag aacctgtact ttcaaagcta cccgtatgac
      661 gttcctgatt atgcctaacc ggcttatcgg tcagtttcac ctgatttacg taaaaacccg
      721 cttcggcggg tttttgcttt tggaggggca gaaagatgaa tgactgtcca cgacgctata
      781 cccaaaagaa aaagcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
      841 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
      901 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
      961 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1021 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1081 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1141 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1201 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1261 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1321 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     1381 gaaattgatc gggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc
     1441 gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     1501 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     1561 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     1621 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     1681 ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat
     1741 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     1801 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     1861 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     1921 cgtttcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     1981 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     2041 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     2101 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttgatatcga
     2161 gctcgcttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt
     2221 tcagaacgct cggttgccgc cgggcgtttt ttattggtca tcgatctcga tcccgcgaaa
     2281 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct agaaataat
//
