LOCUS       sequence-323977-        2606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 nsp6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2606)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2606)
  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-m4046966
FEATURES             Location/Qualifiers
     source          1..2606
                     /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             975..995
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             996..1019
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      1140..1728
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1629..1648
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1834..2493
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1900..1919)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2564..2598
                     /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 gcagtgaaaa
      121 gaacaatcaa gggtacacac cactggttgt tactcacaat tttgacttca cttttagttt
      181 tagtccagag tactcaatgg tctttgttct tttttttgta tgaaaatgcc tttttacctt
      241 ttgctatggg tattattgct atgtctgctt ttgcaatgat gtttgtcaaa cataagcatg
      301 catttctctg tttgtttttg ttaccttctc ttgccactgt agcttatttt aatatggtct
      361 atatgcctgc tagttgggtg atgcgtatta tgacatggtt ggatatggtt gatactagtt
      421 tgtctggttt taagctaaaa gactgtgtta tgtatgcatc agctgtagtg ttactaatcc
      481 ttatgacagc aagaactgtg tatgatgatg gtgctaggag agtgtggaca cttatgaatg
      541 tcttgacact cgtttataaa gtttattatg gtaatgcttt agatcaagcc atttccatgt
      601 gggctcttat aatctctgtt acttctaact actcaggtgt agttacaact gtcatgtttt
      661 tggccagagg tattgttttt atgtgtgttg agtattgccc tattttcttc ataactggta
      721 atacacttca gtgtataatg ctagtttatt gtttcttagg ctatttttgt acttgttact
      781 ttggcctctt ttgtttactc aaccgctact ttagactgac tcttggtgtt tatgattact
      841 tagtttctac acaggagttt agatatatga attcacaggg actactccca cccaagaata
      901 gcatagatgc cttcaaactc aacattaaat tgttgggtgt tggtggcaaa ccttgtatca
      961 aagtagccac tgtagagaac ctgtactttc aaagctggag ccatccgcag tttgaaaaat
     1021 aaccggctta tcggtcagtt tcacctgatt tacgtaaaaa cccgcttcgg cgggtttttg
     1081 cttttggagg ggcagaaaga tgaatgactg tccacgacgc tatacccaaa agaaaaagct
     1141 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1201 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1261 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     1321 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1381 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1441 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1501 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     1561 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     1621 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     1681 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaatt gatcgggcac
     1741 gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag
     1801 ttatcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc
     1861 accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct
     1921 caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag
     1981 aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct
     2041 catccggaat ttcgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac
     2101 ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac
     2161 cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa
     2221 aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtttc agccaatccc
     2281 tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc
     2341 gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt
     2401 caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa
     2461 cagtactgcg atgagtggca gggcggggcg taatttgata tcgagctcgc ttggactcct
     2521 gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg
     2581 ccgccgggcg ttttttattg gtcatc
//
