LOCUS       sequence-287766-        2778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 endoRNAse.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2778)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2778)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046805
FEATURES             Location/Qualifiers
     source          1..2778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             1149..1169
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1170..1199
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     rep_origin      1316..1904
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1805..1824
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2010..2669
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2076..2095)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2740..2774
                     /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 tccttagaaa
      121 atgtcgcctt caacgttgta aacaaaggcc actttgatgg tcagcaaggg gaggttcctg
      181 tgtcaatcat taacaacact gtatacacaa aggtcgacgg cgttgacgtg gaactcttcg
      241 aaaataaaac cactcttccg gttaacgttg catttgagct atgggcgaaa cgtaacatca
      301 aaccggtgcc agaagtaaaa attttgaaca acctgggtgt ggatatcgct gctaatacgg
      361 ttatttggga ctataagaga gatgcgccgg cacatatctc gaccattggt gtttgcagta
      421 tgactgacat cgctaaaaaa cccaccgaaa ccatctgtgc cccgctgacg gtattcttcg
      481 atggccgcgt cgatggtcag gtagacctgt tccggaacgc acgcaatggc gtactgataa
      541 ctgagggatc tgtgaagggc ctgcaaccga gcgttggtcc gaaacaggct agcctgaacg
      601 gtgttaccct gatcggcgaa gcggtcaaaa ctcagtttaa ctactataag aaagtggacg
      661 gtgttgtgca gcagctgccc gaaacgtact tcacacagtc tcgtaacctg caagaattta
      721 aacctcgttc tcagatggaa atcgatttcc tggagctggc tatggacgag tttatcgaac
      781 gctacaaact agaaggctac gccttcgaac acattgtcta tggcgacttc tcccacagcc
      841 agctgggtgg actgcatctg ttgataggtc ttgcgaaacg tttcaaagaa tccccgtttg
      901 agttagaaga ttttattccg atggactcta ctgttaaaaa ttacttcatc accgatgctc
      961 aaaccggcag cagcaaatgc gtttgttctg tcatcgatct gctgctcgat gacttcgtgg
     1021 aaatcattaa gtcccaggat ctgtctgttg tttcaaaagt ggtaaaagta actatcgact
     1081 ataccgagat ttcctttatg ctgtggtgca aggatggcca cgttgaaact ttctacccaa
     1141 aattgcagga gaacctgtac tttcaaagcg agcagaagct tatttccgaa gaagatctct
     1201 aaccggctta tcggtcagtt tcacctgatt tacgtaaaaa cccgcttcgg cgggtttttg
     1261 cttttggagg ggcagaaaga tgaatgactg tccacgacgc tatacccaaa agaaattgag
     1321 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1381 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1441 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     1501 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1561 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1621 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1681 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1741 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     1801 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     1861 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaattgatc gggcacgtaa
     1921 gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat
     1981 cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg
     2041 ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat
     2101 gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa
     2161 ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc
     2221 cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt
     2281 gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg
     2341 acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc
     2401 tggcctattt ccctaaaggg tttattgaga atatgttttt cgtttcagcc aatccctggg
     2461 tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt
     2521 tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg
     2581 ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt
     2641 actgcgatga gtggcagggc ggggcgtaat ttgatatcga gctcgcttgg actcctgttg
     2701 atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc
     2761 cgggcgtttt ttattggt
//
