LOCUS       sequence-298372-        2780 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 2780)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2780)
  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-m4046773
FEATURES             Location/Qualifiers
     source          1..2780
                     /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..1193
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      1314..1902
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1803..1822
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2008..2667
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2074..2093)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2738..2772
                     /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 actacaagga tgatgatgac aaataaccgg
     1201 cttatcggtc agtttcacct gatttacgta aaaacccgct tcggcgggtt tttgcttttg
     1261 gaggggcaga aagatgaatg actgtccacg acgctatacc caaaagaaaa agcttgagat
     1321 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1381 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1441 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     1501 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1561 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1621 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1681 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1741 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1801 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     1861 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aattgatcgg gcacgtaaga
     1921 ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg
     1981 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
     2041 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
     2101 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
     2161 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
     2221 gaatttcgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
     2281 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
     2341 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
     2401 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tttcagccaa tccctgggtg
     2461 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     2521 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     2581 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     2641 tgcgatgagt ggcagggcgg ggcgtaattt gatatcgagc tcgcttggac tcctgttgat
     2701 agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg
     2761 ggcgtttttt attggtcatc
//
