LOCUS       sequence-296837-        2558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SARS-CoV-2 ORF3a protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2558)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2558)
  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-m4046934
FEATURES             Location/Qualifiers
     source          1..2558
                     /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             937..957
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             958..981
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      1102..1690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1591..1610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1796..2455
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1862..1881)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 catcgatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
       61 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagactcttc tatggacttg
      121 tttatgcgta ttttcacaat cggtacggtg accctcaaac agggcgagat caaggatgcg
      181 accccaagtg acttcgttcg cgctactgcc actattccca tccaagcatc tctgcctttt
      241 gggtggctta tcgtaggtgt tgctctgctg gcagtcttcc agtcggcgag caaaataatc
      301 accctaaaaa agcgttggca gctggctctg tctaaaggcg tacatttcgt ctgcaacctg
      361 cttttactgt tcgttactgt gtattcccac ctgttgctgg ttgccgctgg actggaagca
      421 ccgtttctgt acttatacgc tctggtttat ttcttgcaga gcatcaattt tgtgcggata
      481 attatgcgcc tgtggctgtg ttggaaatgc cgttcaaaaa acccgctgct ttacgatgcg
      541 aactacttct tatgctggca caccaactgt tacgattatt gcattccgta taattccgta
      601 accagctcta tcgttatcac gagcggcgac ggcactactt ccccaatcag tgaacatgac
      661 taccagattg gtggttacac tgaaaaatgg gagtctggcg ttaaagattg tgtggtactg
      721 cactcctatt tcacgtctga ctactaccaa ctgtacagca cccagctgtc gaccgacaca
      781 ggtgtcgaac atgtcacttt tttcatctac aacaagatcg ttgatgaacc ggaggaacac
      841 gttcagattc acaccattga tggatcatct ggtgtggtta acccggtgat ggaaccgatc
      901 tatgacgaac ccacgacaac cacttctgta cctctggaga acctgtactt tcaaagcgac
      961 tacaaggatg atgatgacaa ataaccggct tatcggtcag tttcacctga tttacgtaaa
     1021 aacccgcttc ggcgggtttt tgcttttgga ggggcagaaa gatgaatgac tgtccacgac
     1081 gctataccca aaagaaaaag cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1141 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1201 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1261 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1321 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1381 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1441 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1501 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1561 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1621 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1681 cctatggaaa ttgatcgggc acgtaagagg ttccaacttt caccataatg aaataagatc
     1741 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga
     1801 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     1861 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
     1921 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     1981 tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct
     2041 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     2101 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     2161 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     2221 gtttttcgtt tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     2281 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     2341 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     2401 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaatttga
     2461 tatcgagctc gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg
     2521 atttgttcag aacgctcggt tgccgctttt ttattggt
//
