LOCUS       sequence-292488-        2270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 leader protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2270)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2270)
  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-m4046797
FEATURES             Location/Qualifiers
     source          1..2270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    43..67
                     /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             82..104
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             649..669
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             670..687
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      808..1396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1297..1316
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1502..2161
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1568..1587)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2232..2266
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 catcgatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
       61 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagactcttc tatggagagc
      121 cttgtccctg gtttcaacga gaaaacacac gtccaactca gtttgcctgt tttacaggtt
      181 cgcgacgtgc tcgtacgtgg ctttggagac tccgtggagg aggtcttatc agaggcacgt
      241 caacatctta aagatggcac ttgtggctta gtagaagttg aaaaaggcgt tttgcctcaa
      301 cttgaacagc cctatgtgtt catcaaacgt tcggatgctc gaactgcacc tcatggtcat
      361 gttatggttg agctggtagc agaactcgaa ggcattcagt acggtcgtag tggtgagaca
      421 cttggtgtcc ttgtccctca tgtgggcgaa ataccagtgg cttaccgcaa ggttcttctt
      481 cgtaagaacg gtaataaagg agctggtggc catagttacg gcgccgatct aaagtcattt
      541 gacttaggcg acgagcttgg cactgatcct tatgaagatt ttcaagaaaa ctggaacact
      601 aaacatagca gtggtgttac ccgtgaactc atgcgtgagc ttaacggaga gaacctgtac
      661 tttcaaagcc accatcatca ccatcattaa ccggcttatc ggtcagtttc acctgattta
      721 cgtaaaaacc cgcttcggcg ggtttttgct tttggagggg cagaaagatg aatgactgtc
      781 cacgacgcta tacccaaaag aaaaagcttg agatcctttt tttctgcgcg taatctgctg
      841 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
      901 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
      961 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1021 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1081 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1141 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1201 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1261 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1321 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1381 gcggagccta tggaaattga tcgggcacgt aagaggttcc aactttcacc ataatgaaat
     1441 aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa
     1501 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     1561 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     1621 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     1681 tcacattctt gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg
     1741 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     1801 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     1861 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     1921 gaatatgttt ttcgtttcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     1981 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     2041 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     2101 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     2161 atttgatatc gagctcgctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
     2221 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
//
