LOCUS       sequence-292490-        2153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2153)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2153)
  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-m4046871
FEATURES             Location/Qualifiers
     source          1..2153
                     /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             532..552
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             553..570
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      691..1279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1180..1199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1385..2044
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1451..1470)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2115..2149
                     /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 tatggccggt
      121 aacgcaacgg aagttcccgc gaatagcaca gtgctgtctt tttgcgcttt cgctgtagac
      181 gcggcaaaag cttacaagga ttatttggcc tcaggcggac agccaattac caactgtgtc
      241 aaaatgctct gcactcacac cgggactggc caagctataa ccgtaactcc tgaggcaaac
      301 atggaccagg aatccttcgg tggtgcgagt tgctgcctgt actgtcgctg ccatatcgat
      361 cacccgaacc cgaaaggctt ttgtgacctg aaggggaaat acgttcagat cccgactacg
      421 tgcgctaacg atccggttgg tttcaccctg aaaaacaccg tgtgcaccgt gtgcggtatg
      481 tggaaaggct atggctgttc ttgtgaccag ctgagggaac cgatgcttca agagaacctg
      541 tactttcaaa gccaccatca tcaccatcat taaccggctt atcggtcagt ttcacctgat
      601 ttacgtaaaa acccgcttcg gcgggttttt gcttttggag gggcagaaag atgaatgact
      661 gtccacgacg ctatacccaa aagaaaaagc ttgagatcct ttttttctgc gcgtaatctg
      721 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
      781 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
      841 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
      901 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
      961 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1021 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1081 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1141 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     1201 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     1261 ggggcggagc ctatggaaat tgatcgggca cgtaagaggt tccaactttc accataatga
     1321 aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc
     1381 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
     1441 agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct
     1501 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     1561 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga
     1621 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     1681 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     1741 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     1801 tgagaatatg tttttcgttt cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     1861 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     1921 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt
     1981 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc
     2041 gtaatttgat atcgagctcg cttggactcc tgttgataga tccagtaatg acctcagaac
     2101 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggt
//
