LOCUS       sequence-292504-        4526 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 4526)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4526)
  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-m4046799
FEATURES             Location/Qualifiers
     source          1..4526
                     /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             2905..2925
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2926..2943
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      3064..3652
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3553..3572
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3758..4417
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3824..3843)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4488..4522
                     /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 tatggctgat
      121 gcacaatcgt ttttaaaccg ggtttgcggt gtaagtgcag cccgtcttac accgtgcggc
      181 acaggcacta gtactgatgt cgtatacagg gcttttgaca tctacaatga taaagtagct
      241 ggttttgcta aattcctaaa aactaattgt tgtcgcttcc aagaaaagga cgaagatgac
      301 aatttaattg attcttactt tgtagttaag agacacactt tctctaacta ccaacatgaa
      361 gaaacaattt ataatttact taaggattgt ccagctgttg ctaaacatga cttctttaag
      421 tttagaatag acggtgacat ggtaccacat atatcacgtc aacgtcttac taaatacaca
      481 atggcagacc tcgtctatgc tttaaggcat tttgatgaag gtaattgtga cacattaaaa
      541 gaaatacttg tcacatacaa ttgttgtgat gatgattatt tcaataaaaa ggactggtat
      601 gattttgtag aaaacccaga tatattacgc gtatacgcca acttaggtga acgtgtacgc
      661 caagctttgt taaaaacagt acaattctgt gatgccatgc gaaatgctgg tattgttggt
      721 gtactgacat tagataatca agatctcaat ggtaactggt atgatttcgg tgatttcata
      781 caaaccacgc caggtagtgg agttcctgtt gtagattctt attattcatt gttaatgcct
      841 atattaacct tgaccagggc tttaactgca gagtcacatg ttgacactga cttaacaaag
      901 ccttacatta agtgggattt gttaaaatat gacttcacgg aagagaggtt aaaactcttt
      961 gaccgttatt ttaaatattg ggatcagaca taccacccaa attgtgttaa ctgtttggat
     1021 gacagatgca ttctgcattg tgcaaacttt aatgttttat tctctacagt gttcccacct
     1081 acaagttttg gaccactagt gagaaaaata tttgttgatg gtgttccatt tgtagtttca
     1141 actggatacc acttcagaga gctaggtgtt gtacataatc aggatgtaaa cttacatagc
     1201 tctagactta gttttaagga attacttgtg tatgctgctg accctgctat gcacgctgct
     1261 tctggtaatc tattactaga taaacgcact acgtgctttt cagtagctgc acttactaac
     1321 aatgttgctt ttcaaactgt caaacccggt aattttaaca aagacttcta tgactttgct
     1381 gtgtctaagg gtttctttaa ggaaggaagt tctgttgaat taaaacactt cttctttgct
     1441 caggatggta atgctgctat cagcgattat gactactatc gttataatct accaacaatg
     1501 tgtgatatca gacaactact atttgtagtt gaagttgttg ataagtactt tgattgttac
     1561 gatggtggct gtattaatgc taaccaagtc atcgtcaaca acctagacaa atcagctggt
     1621 tttccattta ataaatgggg taaggctaga ctttattatg attcaatgag ttatgaggat
     1681 caagatgcac ttttcgcata tacaaaacgt aatgtcatcc ctactataac tcaaatgaat
     1741 cttaagtatg ccattagtgc aaagaataga gctcgcaccg tagctggtgt ctctatctgt
     1801 agtactatga ccaatagaca gtttcatcaa aaattattga aatcaatagc cgccactaga
     1861 ggagctactg tagtaattgg aacaagcaaa ttctatggtg gttggcacaa catgttaaaa
     1921 actgtttata gtgatgtaga aaaccctcac cttatgggtt gggattatcc taaatgtgat
     1981 agagccatgc ctaacatgct tagaattatg gcctcacttg ttcttgctcg caaacataca
     2041 acgtgttgta gcttgtcaca ccgtttctat agattagcta atgagtgtgc tcaagtattg
     2101 agtgaaatgg tcatgtgtgg cggttcacta tatgttaaac caggtggaac ctcatcagga
     2161 gatgccacaa ctgcttatgc taatagtgtt tttaacattt gtcaagctgt cacggccaat
     2221 gttaatgcac ttttatctac tgatggtaac aaaattgccg ataagtatgt ccgcaattta
     2281 caacacagac tttatgagtg tctctataga aatagagatg ttgacacaga ctttgtgaat
     2341 gagttttacg catatttgcg taaacatttc tcaatgatga tactctctga cgatgctgtt
     2401 gtgtgtttca atagcactta tgcatctcaa ggtctagtgg ctagcataaa gaactttaag
     2461 tcagttcttt attatcaaaa caatgttttt atgtctgaag caaaatgttg gactgagact
     2521 gaccttacta aaggacctca tgaattttgc tctcaacata caatgctagt taaacagggt
     2581 gatgattatg tgtaccttcc ttacccagat ccatcaagaa tcctaggggc cggctgtttt
     2641 gtagatgata tcgtaaaaac agatggtaca cttatgattg aacggttcgt gtctttagct
     2701 atagatgctt acccacttac taaacatcct aatcaggagt atgctgatgt ctttcatttg
     2761 tacttacaat acataagaaa gctacatgat gagttaacag gacacatgtt agacatgtat
     2821 tctgttatgc ttactaatga taacacttca aggtattggg aacctgagtt ttatgaggct
     2881 atgtacacac cgcatacagt cttagagaac ctgtactttc aaagccacca tcatcaccat
     2941 cattaaccgg cttatcggtc agtttcacct gatttacgta aaaacccgct tcggcgggtt
     3001 tttgcttttg gaggggcaga aagatgaatg actgtccacg acgctatacc caaaagaaaa
     3061 agcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3121 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3181 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3241 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3301 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3361 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3421 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3481 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3541 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3601 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aattgatcgg
     3661 gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt
     3721 tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
     3781 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
     3841 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
     3901 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     3961 tgctcatccg gaatttcgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
     4021 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     4081 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     4141 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tttcagccaa
     4201 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     4261 ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
     4321 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
     4381 acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt gatatcgagc tcgcttggac
     4441 tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg
     4501 gttgccgccg ggcgtttttt attggt
//
