LOCUS       sequence-319576-        2564 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 2564)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2564)
  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-m4046856
FEATURES             Location/Qualifiers
     source          1..2564
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             914..934
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      1079..1667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1568..1587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1773..2432
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1839..1858)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2503..2537
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag actcttctat ggatttgttt atgagaatct tcacaattgg
      121 aactgtaact ttgaagcaag gtgaaatcaa ggatgctact ccttcagatt ttgttcgcgc
      181 tactgcaacg ataccgatac aagcctcact ccctttcgga tggcttattg ttggcgttgc
      241 acttcttgct gtttttcaga gcgcttccaa aatcataacc ctcaaaaaga gatggcaact
      301 agcactctcc aagggtgttc actttgtttg caacttgctg ttgttgtttg taacagttta
      361 ctcacacctt ttgctcgttg ctgctggcct tgaagcccct tttctctatc tttatgcttt
      421 agtctacttc ttgcagagta taaactttgt aagaataata atgaggcttt ggctttgctg
      481 gaaatgccgt tccaaaaacc cattacttta tgatgccaac tattttcttt gctggcatac
      541 taattgttac gactattgta taccttacaa tagtgtaact tcttcaattg tcattacttc
      601 aggtgatggc acaacaagtc ctatttctga acatgactac cagattggtg gttatactga
      661 aaaatgggaa tctggagtaa aagactgtgt tgtattacac agttacttca cttcagacta
      721 ttaccagctg tactcaactc aattgagtac agacactggt gttgaacatg ttaccttctt
      781 catctacaat aaaattgttg atgagcctga agaacatgtc caaattcaca caatcgacgg
      841 ttcatccgga gttgttaatc cagtaatgga accaatttat gatgaaccga cgacgactac
      901 tagcgtgcct ttggagaacc tgtactttca aagcgactac aaggatgatg atgacaaata
      961 accggcttat cggtcagttt cacctgattt acgtaaaaac ccgcttcggc gggtttttgc
     1021 ttttggaggg gcagaaagat gaatgactgt ccacgacgct atacccaaaa gaaaaagctt
     1081 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1141 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1201 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     1261 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1321 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1381 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     1441 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     1501 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     1561 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     1621 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaattg atcgggcacg
     1681 taagaggttc caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt
     1741 tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
     1801 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     1861 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     1921 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     1981 atccggaatt tcgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc
     2041 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc
     2101 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa
     2161 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtttca gccaatccct
     2221 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg
     2281 ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc
     2341 aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac
     2401 agtactgcga tgagtggcag ggcggggcgt aatttgatat cgagctcgct tggactcctg
     2461 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
     2521 cgccgggcgt tttttattgg tcatcgatct cgatcccgcg aaat
//
