LOCUS       sequence-298347-        3323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 3'-to-5' exonuclease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3323)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 3323)
  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-m4046878
FEATURES             Location/Qualifiers
     source          1..3323
                     /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             1692..1712
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1713..1736
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      1857..2445
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2346..2365
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2551..3210
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2617..2636)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3281..3315
                     /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 gctgagaatg
      121 tcacgggttt atttaaggac tgttcgaaag tgatcactgg gcttcaccca acccaggccc
      181 ctactcatct gagtgttgat accaaattca aaaccgaagg cctgtgcgtt gacattcccg
      241 gcatcccgaa agatatgact tataggcggc tcatatctat gatgggtttc aagatgaact
      301 accaagtaaa cggctacccg aacatgttca tcacacgtga agaagcgatt cgtcacgtac
      361 gcgcatggat cggttttgac gttgaaggat gccacgctac ccgtgaggcg gtgggtacta
      421 atctgccgct gcaactggga ttctctaccg gcgttaacct ggtcgctgtt ccgacagggt
      481 atgttgatac tccaaacaac acggactttt cacgtgtaag cgctaaaccg ccgccgggcg
      541 atcagttcaa acatctaatc cctttgatgt acaaaggcct gccgtggaac gtggtccgaa
      601 ttaagatcgt gcagatgctt tccgataccc tcaaaaacct gagcgaccgc gtagtattcg
      661 ttctgtgggc ccatggtttt gaactgactt ccatgaagta cttcgtgaaa ataggtccag
      721 agcgcacctg ttgcctttgc gatcgtcgcg caacctgttt tagcactgct tcggacacgt
      781 atgcgtgctg gcaccactct attggcttcg actacgtcta caatcctttc atgatcgacg
      841 ttcagcagtg gggcttcacc ggtaacctgc aatctaacca cgatctgtat tgccaggttc
      901 acggtaacgc tcatgtcgcg tcttgcgatg caattatgac tcgttgcctg gctgtgcacg
      961 aatgcttcgt taaacgcgta gactggacca tcgaatatcc gatcatcggt gatgaactga
     1021 aaatcaatgc agcctgtcgg aaagtacagc atatggttgt taaggctgct ctgctggcag
     1081 acaaatttcc ggtgctgcat gacattggca accctaaagc tatcaaatgc gttccccagg
     1141 cggatgtaga atggaaattc tacgacgcgc aaccatgctc cgataaagcc tacaagattg
     1201 aagaactgtt ctattcatac gcaacacaca gcgacaaatt tactgacggg gtgtgtctgt
     1261 tctggaactg taacgttgac cgttacccgg cgaatagcat cgtctgccgc tttgatacgc
     1321 gtgttttgtc caacctgaac ttaccgggtt gcgatggtgg ctccctctac gtgaacaaac
     1381 acgcattcca cacgcccgct ttcgacaaaa gtgcttttgt gaacctgaag cagctgccgt
     1441 tcttctatta ctctgatagc ccgtgtgagt ctcacggcaa gcaggtagtt tcggatatcg
     1501 actacgttcc gctgaaaagc gccacctgta ttactcgttg caacctcggt ggcgctgttt
     1561 gccgtcacca cgcaaacgag taccgcctgt acctggatgc ttataatatg atgatctccg
     1621 ctggcttttc cttatgggtt tacaaacagt tcgacaccta taacctgtgg aacactttca
     1681 cccgtctgca agagaacctg tactttcaaa gctggagcca tccgcagttt gaaaaataac
     1741 cggcttatcg gtcagtttca cctgatttac gtaaaaaccc gcttcggcgg gtttttgctt
     1801 ttggaggggc agaaagatga atgactgtcc acgacgctat acccaaaaga aaaagcttga
     1861 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1921 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1981 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     2041 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2101 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2161 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2221 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2281 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2341 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2401 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaattgat cgggcacgta
     2461 agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta
     2521 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
     2581 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
     2641 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
     2701 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
     2761 ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
     2821 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
     2881 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
     2941 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtttcagc caatccctgg
     3001 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     3061 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     3121 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     3181 tactgcgatg agtggcaggg cggggcgtaa tttgatatcg agctcgcttg gactcctgtt
     3241 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg
     3301 ccgggcgttt tttattggtc atc
//
