LOCUS       sequence-292520-        3317 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 3317)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 3317)
  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-m4046921
FEATURES             Location/Qualifiers
     source          1..3317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    43..67
                     /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             82..104
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1696..1716
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1717..1734
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      1855..2443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2344..2363
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2549..3208
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2615..2634)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3279..3313
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 catcgatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
       61 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagactcttc tatggctgag
      121 aatgtcacgg gtttatttaa ggactgttcg aaagtgatca ctgggcttca cccaacccag
      181 gcccctactc atctgagtgt tgataccaaa ttcaaaaccg aaggcctgtg cgttgacatt
      241 cccggcatcc cgaaagatat gacttatagg cggctcatat ctatgatggg tttcaagatg
      301 aactaccaag taaacggcta cccgaacatg ttcatcacac gtgaagaagc gattcgtcac
      361 gtacgcgcat ggatcggttt tgacgttgaa ggatgccacg ctacccgtga ggcggtgggt
      421 actaatctgc cgctgcaact gggattctct accggcgtta acctggtcgc tgttccgaca
      481 gggtatgttg atactccaaa caacacggac ttttcacgtg taagcgctaa accgccgccg
      541 ggcgatcagt tcaaacatct aatccctttg atgtacaaag gcctgccgtg gaacgtggtc
      601 cgaattaaga tcgtgcagat gctttccgat accctcaaaa acctgagcga ccgcgtagta
      661 ttcgttctgt gggcccatgg ttttgaactg acttccatga agtacttcgt gaaaataggt
      721 ccagagcgca cctgttgcct ttgcgatcgt cgcgcaacct gttttagcac tgcttcggac
      781 acgtatgcgt gctggcacca ctctattggc ttcgactacg tctacaatcc tttcatgatc
      841 gacgttcagc agtggggctt caccggtaac ctgcaatcta accacgatct gtattgccag
      901 gttcacggta acgctcatgt cgcgtcttgc gatgcaatta tgactcgttg cctggctgtg
      961 cacgaatgct tcgttaaacg cgtagactgg accatcgaat atccgatcat cggtgatgaa
     1021 ctgaaaatca atgcagcctg tcggaaagta cagcatatgg ttgttaaggc tgctctgctg
     1081 gcagacaaat ttccggtgct gcatgacatt ggcaacccta aagctatcaa atgcgttccc
     1141 caggcggatg tagaatggaa attctacgac gcgcaaccat gctccgataa agcctacaag
     1201 attgaagaac tgttctattc atacgcaaca cacagcgaca aatttactga cggggtgtgt
     1261 ctgttctgga actgtaacgt tgaccgttac ccggcgaata gcatcgtctg ccgctttgat
     1321 acgcgtgttt tgtccaacct gaacttaccg ggttgcgatg gtggctccct ctacgtgaac
     1381 aaacacgcat tccacacgcc cgctttcgac aaaagtgctt ttgtgaacct gaagcagctg
     1441 ccgttcttct attactctga tagcccgtgt gagtctcacg gcaagcaggt agtttcggat
     1501 atcgactacg ttccgctgaa aagcgccacc tgtattactc gttgcaacct cggtggcgct
     1561 gtttgccgtc accacgcaaa cgagtaccgc ctgtacctgg atgcttataa tatgatgatc
     1621 tccgctggct tttccttatg ggtttacaaa cagttcgaca cctataacct gtggaacact
     1681 ttcacccgtc tgcaagagaa cctgtacttt caaagccacc atcatcacca tcattaaccg
     1741 gcttatcggt cagtttcacc tgatttacgt aaaaacccgc ttcggcgggt ttttgctttt
     1801 ggaggggcag aaagatgaat gactgtccac gacgctatac ccaaaagaaa aagcttgaga
     1861 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     1921 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     1981 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     2041 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2101 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2161 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2221 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2281 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2341 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2401 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaattgatcg ggcacgtaag
     2461 aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc
     2521 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
     2581 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
     2641 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
     2701 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
     2761 ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
     2821 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
     2881 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     2941 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtttcagcca atccctgggt
     3001 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     3061 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     3121 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     3181 ctgcgatgag tggcagggcg gggcgtaatt tgatatcgag ctcgcttgga ctcctgttga
     3241 tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc
     3301 gggcgttttt tattggt
//
