LOCUS       sequence-292502-        1985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1985)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1985)
  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-m4046979
FEATURES             Location/Qualifiers
     source          1..1985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      139..173
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     primer_bind     201..220
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        201..219
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    220..244
                     /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             259..281
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             541..561
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             562..579
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      700..1288
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1189..1208
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(1394..1985,1..68)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1460..1479)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
       61 gggcgtaatt tgatatcgag ctcgcttgga ctcctgttga tagatccagt aatgacctca
      121 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtcat
      181 cgatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
      241 ttcccctcta gaaataattt tgtttaactt taagaaggag actcttctat gagcaaaatg
      301 tctgatgtga agtgtacttc cgttgtattg ctgtcagtcc tgcaacagct gcgtgttgag
      361 tcgtctagca aactatgggc acagtgcgtg cagctgcaca acgacatcct cttagcgaaa
      421 gatacgaccg aagctttcga aaaaatggtt tcgctgttgt ccgtacttct gtctatgcaa
      481 ggtgccgttg acattaataa actgtgcgaa gagatgctgg acaaccgcgc tactctgcaa
      541 gagaacctgt actttcaaag ccaccatcat caccatcatt aaccggctta tcggtcagtt
      601 tcacctgatt tacgtaaaaa cccgcttcgg cgggtttttg cttttggagg ggcagaaaga
      661 tgaatgactg tccacgacgc tatacccaaa agaaaaagct tgagatcctt tttttctgcg
      721 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
      781 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      841 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      901 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
      961 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1021 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1081 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1141 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1201 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1261 ctcgtcaggg gggcggagcc tatggaaatt gatcgggcac gtaagaggtt ccaactttca
     1321 ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc
     1381 taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg
     1441 gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac
     1501 cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta
     1561 tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat ttcgtatggc
     1621 aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca
     1681 tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt
     1741 tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa
     1801 agggtttatt gagaatatgt ttttcgtttc agccaatccc tgggtgagtt tcaccagttt
     1861 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata
     1921 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg
     1981 tgatg
//
