LOCUS       sequence-297847-        2273 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 2273)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2273)
  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-m4046854
FEATURES             Location/Qualifiers
     source          1..2273
                     /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             648..668
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             669..686
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      807..1395
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1296..1315
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1501..2160
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1567..1586)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2231..2265
                     /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 gagtctctag
      121 tgcctgggtt caacgaaaaa actcatgtcc agctgtcgtt accagttctg caagtacgcg
      181 atgttctcgt tcgtggtttt ggagacagcg tagaagaagt tttgagtgaa gcccgtcagc
      241 acctgaagga cggcacctgt ggcctggtgg aggtggaaaa aggtgtcctt ccgcagctgg
      301 aacagcccta tgttttcatt aaacgatccg atgctcgcac ggcgccgcac ggtcacgtca
      361 tggtagagct ggttgcagaa ttggaaggca tccagtacgg tcgttctggc gaaactctgg
      421 gggtacttgt gccgcatgtt ggtgagatcc cggttgctta ccgtaaagtg ctgctgcgga
      481 agaatggcaa caaaggcgcg ggtggtcaca gctatggcgc tgatctgaaa tcattcgacc
      541 tgggagatga actgggtacc gacccgtacg aggattttca agaaaactgg aacacaaaac
      601 attctagcgg cgttactcgt gagctgatga gggaactcaa tggtggcgag aacctgtact
      661 ttcaaagcca ccatcatcac catcattaac cggcttatcg gtcagtttca cctgatttac
      721 gtaaaaaccc gcttcggcgg gtttttgctt ttggaggggc agaaagatga atgactgtcc
      781 acgacgctat acccaaaaga aaaagcttga gatccttttt ttctgcgcgt aatctgctgc
      841 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
      901 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
      961 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     1021 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     1081 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     1141 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     1201 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     1261 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     1321 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     1381 cggagcctat ggaaattgat cgggcacgta agaggttcca actttcacca taatgaaata
     1441 agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa
     1501 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
     1561 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat
     1621 attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt
     1681 cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt
     1741 gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa
     1801 acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat
     1861 tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag
     1921 aatatgtttt tcgtttcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg
     1981 gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc
     2041 gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat
     2101 gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa
     2161 tttgatatcg agctcgcttg gactcctgtt gatagatcca gtaatgacct cagaactcca
     2221 tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtc atc
//
