LOCUS       sequence-287808-        2274 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 2274)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2274)
  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-m4046821
FEATURES             Location/Qualifiers
     source          1..2274
                     /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             645..665
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             666..695
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     rep_origin      812..1400
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1301..1320
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1506..2165
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1572..1591)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2236..2270
                     /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 gagagccttg
      121 tccctggttt caacgagaaa acacacgtcc aactcagttt gcctgtttta caggttcgcg
      181 acgtgctcgt acgtggcttt ggagactccg tggaggaggt cttatcagag gcacgtcaac
      241 atcttaaaga tggcacttgt ggcttagtag aagttgaaaa aggcgttttg cctcaacttg
      301 aacagcccta tgtgttcatc aaacgttcgg atgctcgaac tgcacctcat ggtcatgtta
      361 tggttgagct ggtagcagaa ctcgaaggca ttcagtacgg tcgtagtggt gagacacttg
      421 gtgtccttgt ccctcatgtg ggcgaaatac cagtggctta ccgcaaggtt cttcttcgta
      481 agaacggtaa taaaggagct ggtggccata gttacggcgc cgatctaaag tcatttgact
      541 taggcgacga gcttggcact gatccttatg aagattttca agaaaactgg aacactaaac
      601 atagcagtgg tgttacccgt gaactcatgc gtgagcttaa cggagagaac ctgtactttc
      661 aaagcgagca gaagcttatt tccgaagaag atctctaacc ggcttatcgg tcagtttcac
      721 ctgatttacg taaaaacccg cttcggcggg tttttgcttt tggaggggca gaaagatgaa
      781 tgactgtcca cgacgctata cccaaaagaa attgagatcc tttttttctg cgcgtaatct
      841 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
      901 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
      961 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1021 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1081 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1141 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1201 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1261 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1321 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1381 gggggcggag cctatggaaa ttgatcgggc acgtaagagg ttccaacttt caccataatg
     1441 aaataagatc actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag
     1501 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     1561 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     1621 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     1681 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag
     1741 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     1801 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     1861 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     1921 ttgagaatat gtttttcgtt tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     1981 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     2041 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     2101 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     2161 cgtaatttga tatcgagctc gcttggactc ctgttgatag atccagtaat gacctcagaa
     2221 ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat tggt
//
