LOCUS       sequence-287807-        2271 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 2271)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2271)
  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-m4046782
FEATURES             Location/Qualifiers
     source          1..2271
                     /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..692
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      809..1397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1298..1317
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1503..2162
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1569..1588)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2233..2267
                     /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 aaagctaccc gtatgacgtt cctgattatg cctaaccggc ttatcggtca gtttcacctg
      721 atttacgtaa aaacccgctt cggcgggttt ttgcttttgg aggggcagaa agatgaatga
      781 ctgtccacga cgctataccc aaaagaaatt gagatccttt ttttctgcgc gtaatctgct
      841 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
      901 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
      961 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1021 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1081 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1141 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1201 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1261 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1321 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1381 ggcggagcct atggaaattg atcgggcacg taagaggttc caactttcac cataatgaaa
     1441 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
     1501 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     1561 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     1621 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     1681 ttcacattct tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg
     1741 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     1801 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     1861 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     1921 agaatatgtt tttcgtttca gccaatccct gggtgagttt caccagtttt gatttaaacg
     1981 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     2041 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     2101 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     2161 aatttgatat cgagctcgct tggactcctg ttgatagatc cagtaatgac ctcagaactc
     2221 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg t
//
