LOCUS       sequence-287797-        1860 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 ORF7b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1860)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1860)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046917
FEATURES             Location/Qualifiers
     source          1..1860
                     /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             237..257
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             258..281
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      398..986
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     887..906
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1092..1751
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1158..1177)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1822..1856
                     /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 attgaattgt
      121 cactgatcga tttctatctt tgttttctcg cgttcctgct gtttctggtt ctgatcatgc
      181 tgattatctt ctggttctcc ttagagctgc aagaccataa tgaaacctgc cacgccgaga
      241 acctgtactt tcaaagctgg agccatccgc agtttgaaaa ataaccggct tatcggtcag
      301 tttcacctga tttacgtaaa aacccgcttc ggcgggtttt tgcttttgga ggggcagaaa
      361 gatgaatgac tgtccacgac gctataccca aaagaaattg agatcctttt tttctgcgcg
      421 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
      481 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
      541 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
      601 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
      661 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
      721 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
      781 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
      841 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
      901 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
      961 cgtcaggggg gcggagccta tggaaattga tcgggcacgt aagaggttcc aactttcacc
     1021 ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta
     1081 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     1141 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     1201 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     1261 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattt cgtatggcaa
     1321 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     1381 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     1441 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     1501 ggtttattga gaatatgttt ttcgtttcag ccaatccctg ggtgagtttc accagttttg
     1561 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
     1621 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
     1681 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     1741 gcggggcgta atttgatatc gagctcgctt ggactcctgt tgatagatcc agtaatgacc
     1801 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
//
