LOCUS       sequence-297842-        4535 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 RNA-dependent RNA 
            polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4535)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4535)
  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-m4046802
FEATURES             Location/Qualifiers
     source          1..4535
                     /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             2901..2921
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2922..2948
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      3069..3657
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3558..3577
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3763..4422
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3829..3848)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4493..4527
                     /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 aaaccgggtt tgcggtgtaa gtgcagcccg tcttacaccg tgcggcacag
      181 gcactagtac tgatgtcgta tacagggctt ttgacatcta caatgataaa gtagctggtt
      241 ttgctaaatt cctaaaaact aattgttgtc gcttccaaga aaaggacgaa gatgacaatt
      301 taattgattc ttactttgta gttaagagac acactttctc taactaccaa catgaagaaa
      361 caatttataa tttacttaag gattgtccag ctgttgctaa acatgacttc tttaagttta
      421 gaatagacgg tgacatggta ccacatatat cacgtcaacg tcttactaaa tacacaatgg
      481 cagacctcgt ctatgcttta aggcattttg atgaaggtaa ttgtgacaca ttaaaagaaa
      541 tacttgtcac atacaattgt tgtgatgatg attatttcaa taaaaaggac tggtatgatt
      601 ttgtagaaaa cccagatata ttacgcgtat acgccaactt aggtgaacgt gtacgccaag
      661 ctttgttaaa aacagtacaa ttctgtgatg ccatgcgaaa tgctggtatt gttggtgtac
      721 tgacattaga taatcaagat ctcaatggta actggtatga tttcggtgat ttcatacaaa
      781 ccacgccagg tagtggagtt cctgttgtag attcttatta ttcattgtta atgcctatat
      841 taaccttgac cagggcttta actgcagagt cacatgttga cactgactta acaaagcctt
      901 acattaagtg ggatttgtta aaatatgact tcacggaaga gaggttaaaa ctctttgacc
      961 gttattttaa atattgggat cagacatacc acccaaattg tgttaactgt ttggatgaca
     1021 gatgcattct gcattgtgca aactttaatg ttttattctc tacagtgttc ccacctacaa
     1081 gttttggacc actagtgaga aaaatatttg ttgatggtgt tccatttgta gtttcaactg
     1141 gataccactt cagagagcta ggtgttgtac ataatcagga tgtaaactta catagctcta
     1201 gacttagttt taaggaatta cttgtgtatg ctgctgaccc tgctatgcac gctgcttctg
     1261 gtaatctatt actagataaa cgcactacgt gcttttcagt agctgcactt actaacaatg
     1321 ttgcttttca aactgtcaaa cccggtaatt ttaacaaaga cttctatgac tttgctgtgt
     1381 ctaagggttt ctttaaggaa ggaagttctg ttgaattaaa acacttcttc tttgctcagg
     1441 atggtaatgc tgctatcagc gattatgact actatcgtta taatctacca acaatgtgtg
     1501 atatcagaca actactattt gtagttgaag ttgttgataa gtactttgat tgttacgatg
     1561 gtggctgtat taatgctaac caagtcatcg tcaacaacct agacaaatca gctggttttc
     1621 catttaataa atggggtaag gctagacttt attatgattc aatgagttat gaggatcaag
     1681 atgcactttt cgcatataca aaacgtaatg tcatccctac tataactcaa atgaatctta
     1741 agtatgccat tagtgcaaag aatagagctc gcaccgtagc tggtgtctct atctgtagta
     1801 ctatgaccaa tagacagttt catcaaaaat tattgaaatc aatagccgcc actagaggag
     1861 ctactgtagt aattggaaca agcaaattct atggtggttg gcacaacatg ttaaaaactg
     1921 tttatagtga tgtagaaaac cctcacctta tgggttggga ttatcctaaa tgtgatagag
     1981 ccatgcctaa catgcttaga attatggcct cacttgttct tgctcgcaaa catacaacgt
     2041 gttgtagctt gtcacaccgt ttctatagat tagctaatga gtgtgctcaa gtattgagtg
     2101 aaatggtcat gtgtggcggt tcactatatg ttaaaccagg tggaacctca tcaggagatg
     2161 ccacaactgc ttatgctaat agtgttttta acatttgtca agctgtcacg gccaatgtta
     2221 atgcactttt atctactgat ggtaacaaaa ttgccgataa gtatgtccgc aatttacaac
     2281 acagacttta tgagtgtctc tatagaaata gagatgttga cacagacttt gtgaatgagt
     2341 tttacgcata tttgcgtaaa catttctcaa tgatgatact ctctgacgat gctgttgtgt
     2401 gtttcaatag cacttatgca tctcaaggtc tagtggctag cataaagaac tttaagtcag
     2461 ttctttatta tcaaaacaat gtttttatgt ctgaagcaaa atgttggact gagactgacc
     2521 ttactaaagg acctcatgaa ttttgctctc aacatacaat gctagttaaa cagggtgatg
     2581 attatgtgta ccttccttac ccagatccat caagaatcct aggggccggc tgttttgtag
     2641 atgatatcgt aaaaacagat ggtacactta tgattgaacg gttcgtgtct ttagctatag
     2701 atgcttaccc acttactaaa catcctaatc aggagtatgc tgatgtcttt catttgtact
     2761 tacaatacat aagaaagcta catgatgagt taacaggaca catgttagac atgtattctg
     2821 ttatgcttac taatgataac acttcaaggt attgggaacc tgagttttat gaggctatgt
     2881 acacaccgca tacagtctta gagaacctgt actttcaaag ctacccgtat gacgttcctg
     2941 attatgccta accggcttat cggtcagttt cacctgattt acgtaaaaac ccgcttcggc
     3001 gggtttttgc ttttggaggg gcagaaagat gaatgactgt ccacgacgct atacccaaaa
     3061 gaaaaagctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3121 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3181 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3241 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3301 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3361 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3421 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3481 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3541 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3601 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaattg
     3661 atcgggcacg taagaggttc caactttcac cataatgaaa taagatcact accgggcgta
     3721 ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact
     3781 ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag
     3841 tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag
     3901 accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg
     3961 atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt gatatgggat
     4021 agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg
     4081 agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt
     4141 tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtttca
     4201 gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc
     4261 ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg
     4321 ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat
     4381 gaattacaac agtactgcga tgagtggcag ggcggggcgt aatttgatat cgagctcgct
     4441 tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac
     4501 gctcggttgc cgccgggcgt tttttattgg tcatc
//
