LOCUS       sequence-287831-        1977 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 1977)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1977)
  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-m4046786
FEATURES             Location/Qualifiers
     source          1..1977
                     /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..398
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      515..1103
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1004..1023
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1209..1868
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1275..1294)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1939..1973
                     /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 aagcgactac aaggatgatg atgacaaata accggcttat cggtcagttt
      421 cacctgattt acgtaaaaac ccgcttcggc gggtttttgc ttttggaggg gcagaaagat
      481 gaatgactgt ccacgacgct atacccaaaa gaaattgaga tccttttttt ctgcgcgtaa
      541 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
      601 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
      661 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
      721 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
      781 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
      841 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
      901 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
      961 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     1021 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     1081 caggggggcg gagcctatgg aaattgatcg ggcacgtaag aggttccaac tttcaccata
     1141 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg
     1201 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     1261 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     1321 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     1381 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaatttcgt atggcaatga
     1441 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     1501 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     1561 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     1621 ttattgagaa tatgtttttc gtttcagcca atccctgggt gagtttcacc agttttgatt
     1681 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
     1741 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     1801 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     1861 gggcgtaatt tgatatcgag ctcgcttgga ctcctgttga tagatccagt aatgacctca
     1921 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggt
//
