LOCUS       Exported                2783 bp ds-DNA     circular SYN 20-JAN-2023
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 endoRNAse.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046838
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2783)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2783)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 20, 2023 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2783
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1130..1150
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1151..1177
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      1298..1886
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1787..1806
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1992..2651
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2058..2077)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2722..2756
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag actcttctat gtccttagaa aatgtcgcct tcaacgttgt
      121 aaacaaaggc cactttgatg gtcagcaagg ggaggttcct gtgtcaatca ttaacaacac
      181 tgtatacaca aaggtcgacg gcgttgacgt ggaactcttc gaaaataaaa ccactcttcc
      241 ggttaacgtt gcatttgagc tatgggcgaa acgtaacatc aaaccggtgc cagaagtaaa
      301 aattttgaac aacctgggtg tggatatcgc tgctaatacg gttatttggg actataagag
      361 agatgcgccg gcacatatct cgaccattgg tgtttgcagt atgactgaca tcgctaaaaa
      421 acccaccgaa accatctgtg ccccgctgac ggtattcttc gatggccgcg tcgatggtca
      481 ggtagacctg ttccggaacg cacgcaatgg cgtactgata actgagggat ctgtgaaggg
      541 cctgcaaccg agcgttggtc cgaaacaggc tagcctgaac ggtgttaccc tgatcggcga
      601 agcggtcaaa actcagttta actactataa gaaagtggac ggtgttgtgc agcagctgcc
      661 cgaaacgtac ttcacacagt ctcgtaacct gcaagaattt aaacctcgtt ctcagatgga
      721 aatcgatttc ctggagctgg ctatggacga gtttatcgaa cgctacaaac tagaaggcta
      781 cgccttcgaa cacattgtct atggcgactt ctcccacagc cagctgggtg gactgcatct
      841 gttgataggt cttgcgaaac gtttcaaaga atccccgttt gagttagaag attttattcc
      901 gatggactct actgttaaaa attacttcat caccgatgct caaaccggca gcagcaaatg
      961 cgtttgttct gtcatcgatc tgctgctcga tgacttcgtg gaaatcatta agtcccagga
     1021 tctgtctgtt gtttcaaaag tggtaaaagt aactatcgac tataccgaga tttcctttat
     1081 gctgtggtgc aaggatggcc acgttgaaac tttctaccca aaattgcagg agaacctgta
     1141 ctttcaaagc tacccgtatg acgttcctga ttatgcctaa ccggcttatc ggtcagtttc
     1201 acctgattta cgtaaaaacc cgcttcggcg ggtttttgct tttggagggg cagaaagatg
     1261 aatgactgtc cacgacgcta tacccaaaag aaaaagcttg agatcctttt tttctgcgcg
     1321 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1381 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1441 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1501 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1561 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     1621 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     1681 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1741 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1801 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     1861 cgtcaggggg gcggagccta tggaaattga tcgggcacgt aagaggttcc aactttcacc
     1921 ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta
     1981 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     2041 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     2101 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     2161 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattt cgtatggcaa
     2221 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     2281 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     2341 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     2401 ggtttattga gaatatgttt ttcgtttcag ccaatccctg ggtgagtttc accagttttg
     2461 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
     2521 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
     2581 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     2641 gcggggcgta atttgatatc gagctcgctt ggactcctgt tgatagatcc agtaatgacc
     2701 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
     2761 catcgatctc gatcccgcga aat
//
