LOCUS       sequence-323992-        2276 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 2276)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2276)
  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-m4046770
FEATURES             Location/Qualifiers
     source          1..2276
                     /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..689
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      810..1398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1299..1318
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1504..2163
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1570..1589)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2234..2268
                     /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 aaagctggag ccatccgcag tttgaaaaat aaccggctta tcggtcagtt tcacctgatt
      721 tacgtaaaaa cccgcttcgg cgggtttttg cttttggagg ggcagaaaga tgaatgactg
      781 tccacgacgc tatacccaaa agaaaaagct tgagatcctt tttttctgcg cgtaatctgc
      841 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
      901 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
      961 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1021 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     1081 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     1141 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     1201 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     1261 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     1321 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     1381 gggcggagcc tatggaaatt gatcgggcac gtaagaggtt ccaactttca ccataatgaa
     1441 ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct
     1501 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
     1561 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
     1621 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt
     1681 attcacattc ttgcccgcct gatgaatgct catccggaat ttcgtatggc aatgaaagac
     1741 ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact
     1801 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata
     1861 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt
     1921 gagaatatgt ttttcgtttc agccaatccc tgggtgagtt tcaccagttt tgatttaaac
     1981 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa
     2041 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc
     2101 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg
     2161 taatttgata tcgagctcgc ttggactcct gttgatagat ccagtaatga cctcagaact
     2221 ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtcatc
//
