LOCUS       sequence-292572-        2774 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 2774)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2774)
  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-m4046740
FEATURES             Location/Qualifiers
     source          1..2774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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..1168
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      1289..1877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1778..1797
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1983..2642
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2049..2068)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2713..2747
                     /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 caccatcatc accatcatta accggcttat cggtcagttt cacctgattt
     1201 acgtaaaaac ccgcttcggc gggtttttgc ttttggaggg gcagaaagat gaatgactgt
     1261 ccacgacgct atacccaaaa gaaaaagctt gagatccttt ttttctgcgc gtaatctgct
     1321 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1381 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1441 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1501 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1561 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1621 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1681 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1741 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1801 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1861 ggcggagcct atggaaattg atcgggcacg taagaggttc caactttcac cataatgaaa
     1921 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
     1981 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     2041 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     2101 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     2161 ttcacattct tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg
     2221 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     2281 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     2341 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     2401 agaatatgtt tttcgtttca gccaatccct gggtgagttt caccagtttt gatttaaacg
     2461 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     2521 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     2581 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     2641 aatttgatat cgagctcgct tggactcctg ttgatagatc cagtaatgac ctcagaactc
     2701 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg tcatcgatct
     2761 cgatcccgcg aaat
//
