LOCUS       sequence-292496-        1862 bp    DNA     circular SYN 15-DEC-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 1862)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1862)
  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-m4046859
FEATURES             Location/Qualifiers
     source          1..1862
                     /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             218..238
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             239..256
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      377..965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     866..885
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1071..1730
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1137..1156)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1801..1835
                     /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 gattgaactt tcattaattg acttctattt
      121 gtgcttttta gcctttctgc tattccttgt tttaattatg cttattatct tttggttctc
      181 acttgaactg caagatcata atgaaacttg tcacgccgag aacctgtact ttcaaagcca
      241 ccatcatcac catcattaac cggcttatcg gtcagtttca cctgatttac gtaaaaaccc
      301 gcttcggcgg gtttttgctt ttggaggggc agaaagatga atgactgtcc acgacgctat
      361 acccaaaaga aaaagcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      421 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      481 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      541 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      601 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      661 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      721 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      781 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      841 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      901 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      961 ggaaattgat cgggcacgta agaggttcca actttcacca taatgaaata agatcactac
     1021 cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa
     1081 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
     1141 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
     1201 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
     1261 cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt gagctggtga
     1321 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
     1381 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
     1441 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
     1501 tcgtttcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
     1561 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc
     1621 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     1681 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttgatatcg
     1741 agctcgcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
     1801 ttcagaacgc tcggttgccg ccgggcgttt tttattggtc atcgatctcg atcccgcgaa
     1861 at
//
