LOCUS       sequence-287753-        2772 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 2772)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2772)
  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-m4046734
FEATURES             Location/Qualifiers
     source          1..2772
                     /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="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      1310..1898
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1799..1818
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2004..2663
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2070..2089)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2734..2768
                     /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 tttcaaagct ggagccatcc gcagtttgaa aaataaccgg
     1201 cttatcggtc agtttcacct gatttacgta aaaacccgct tcggcgggtt tttgcttttg
     1261 gaggggcaga aagatgaatg actgtccacg acgctatacc caaaagaaat tgagatcctt
     1321 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1381 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1441 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     1501 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     1561 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     1621 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     1681 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     1741 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     1801 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     1861 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaatt gatcgggcac gtaagaggtt
     1921 ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat
     1981 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     2041 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     2101 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     2161 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     2221 ttcgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     2281 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     2341 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     2401 atttccctaa agggtttatt gagaatatgt ttttcgtttc agccaatccc tgggtgagtt
     2461 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     2521 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     2581 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     2641 atgagtggca gggcggggcg taatttgata tcgagctcgc ttggactcct gttgatagat
     2701 ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg
     2761 ttttttattg gt
//
