LOCUS       sequence-287769-        1971 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 1971)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1971)
  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-m4046933
FEATURES             Location/Qualifiers
     source          1..1971
                     /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             354..374
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             375..392
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      509..1097
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     998..1017
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1203..1862
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1269..1288)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1933..1967
                     /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 aaaatgtcag
      121 atgtaaagtg cacatcagta gtcttactct cagttttgca acaactcaga gtagaatcat
      181 catctaaatt gtgggctcaa tgtgtccagt tacacaatga cattctctta gctaaagata
      241 ctactgaagc ctttgaaaaa atggtttcac tactttctgt tttgctttcc atgcagggtg
      301 ctgtagacat aaacaagctt tgtgaagaaa tgctggacaa cagggcaacc ttagagaacc
      361 tgtactttca aagccaccat catcaccatc attaaccggc ttatcggtca gtttcacctg
      421 atttacgtaa aaacccgctt cggcgggttt ttgcttttgg aggggcagaa agatgaatga
      481 ctgtccacga cgctataccc aaaagaaatt gagatccttt ttttctgcgc gtaatctgct
      541 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
      601 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
      661 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
      721 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
      781 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
      841 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
      901 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
      961 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1021 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1081 ggcggagcct atggaaattg atcgggcacg taagaggttc caactttcac cataatgaaa
     1141 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
     1201 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     1261 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     1321 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     1381 ttcacattct tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg
     1441 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     1501 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     1561 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     1621 agaatatgtt tttcgtttca gccaatccct gggtgagttt caccagtttt gatttaaacg
     1681 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     1741 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     1801 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     1861 aatttgatat cgagctcgct tggactcctg ttgatagatc cagtaatgac ctcagaactc
     1921 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg t
//
