LOCUS       sequence-287770-        1761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1761)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1761)
  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-m4046974
FEATURES             Location/Qualifiers
     source          1..1761
                     /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             144..164
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             165..182
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      299..887
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     788..807
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             993..1652
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1059..1078)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1723..1757
                     /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 gctgatgcac
      121 aatcgttttt aaacgggttt gcggagaacc tgtactttca aagccaccat catcaccatc
      181 attaaccggc ttatcggtca gtttcacctg atttacgtaa aaacccgctt cggcgggttt
      241 ttgcttttgg aggggcagaa agatgaatga ctgtccacga cgctataccc aaaagaaatt
      301 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
      361 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
      421 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
      481 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
      541 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
      601 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
      661 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
      721 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      781 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      841 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaattg atcgggcacg
      901 taagaggttc caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt
      961 tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
     1021 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     1081 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     1141 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     1201 atccggaatt tcgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc
     1261 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc
     1321 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa
     1381 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtttca gccaatccct
     1441 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg
     1501 ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc
     1561 aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac
     1621 agtactgcga tgagtggcag ggcggggcgt aatttgatat cgagctcgct tggactcctg
     1681 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
     1741 cgccgggcgt tttttattgg t
//
