LOCUS       sequence-319574-        4538 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 4538)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4538)
  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-m4046831
FEATURES             Location/Qualifiers
     source          1..4538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..24
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5..23
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    24..48
                     /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             63..85
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2886..2906
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2907..2936
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     rep_origin      3057..3645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3546..3565
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3751..4410
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3817..3836)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4481..4515
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
       61 attttgttta actttaagaa ggagactctt ctatggctga tgcacaatcg tttttaaacc
      121 gggtttgcgg tgtaagtgca gcccgtctta caccgtgcgg cacaggcact agtactgatg
      181 tcgtatacag ggcttttgac atctacaatg ataaagtagc tggttttgct aaattcctaa
      241 aaactaattg ttgtcgcttc caagaaaagg acgaagatga caatttaatt gattcttact
      301 ttgtagttaa gagacacact ttctctaact accaacatga agaaacaatt tataatttac
      361 ttaaggattg tccagctgtt gctaaacatg acttctttaa gtttagaata gacggtgaca
      421 tggtaccaca tatatcacgt caacgtctta ctaaatacac aatggcagac ctcgtctatg
      481 ctttaaggca ttttgatgaa ggtaattgtg acacattaaa agaaatactt gtcacataca
      541 attgttgtga tgatgattat ttcaataaaa aggactggta tgattttgta gaaaacccag
      601 atatattacg cgtatacgcc aacttaggtg aacgtgtacg ccaagctttg ttaaaaacag
      661 tacaattctg tgatgccatg cgaaatgctg gtattgttgg tgtactgaca ttagataatc
      721 aagatctcaa tggtaactgg tatgatttcg gtgatttcat acaaaccacg ccaggtagtg
      781 gagttcctgt tgtagattct tattattcat tgttaatgcc tatattaacc ttgaccaggg
      841 ctttaactgc agagtcacat gttgacactg acttaacaaa gccttacatt aagtgggatt
      901 tgttaaaata tgacttcacg gaagagaggt taaaactctt tgaccgttat tttaaatatt
      961 gggatcagac ataccaccca aattgtgtta actgtttgga tgacagatgc attctgcatt
     1021 gtgcaaactt taatgtttta ttctctacag tgttcccacc tacaagtttt ggaccactag
     1081 tgagaaaaat atttgttgat ggtgttccat ttgtagtttc aactggatac cacttcagag
     1141 agctaggtgt tgtacataat caggatgtaa acttacatag ctctagactt agttttaagg
     1201 aattacttgt gtatgctgct gaccctgcta tgcacgctgc ttctggtaat ctattactag
     1261 ataaacgcac tacgtgcttt tcagtagctg cacttactaa caatgttgct tttcaaactg
     1321 tcaaacccgg taattttaac aaagacttct atgactttgc tgtgtctaag ggtttcttta
     1381 aggaaggaag ttctgttgaa ttaaaacact tcttctttgc tcaggatggt aatgctgcta
     1441 tcagcgatta tgactactat cgttataatc taccaacaat gtgtgatatc agacaactac
     1501 tatttgtagt tgaagttgtt gataagtact ttgattgtta cgatggtggc tgtattaatg
     1561 ctaaccaagt catcgtcaac aacctagaca aatcagctgg ttttccattt aataaatggg
     1621 gtaaggctag actttattat gattcaatga gttatgagga tcaagatgca cttttcgcat
     1681 atacaaaacg taatgtcatc cctactataa ctcaaatgaa tcttaagtat gccattagtg
     1741 caaagaatag agctcgcacc gtagctggtg tctctatctg tagtactatg accaatagac
     1801 agtttcatca aaaattattg aaatcaatag ccgccactag aggagctact gtagtaattg
     1861 gaacaagcaa attctatggt ggttggcaca acatgttaaa aactgtttat agtgatgtag
     1921 aaaaccctca ccttatgggt tgggattatc ctaaatgtga tagagccatg cctaacatgc
     1981 ttagaattat ggcctcactt gttcttgctc gcaaacatac aacgtgttgt agcttgtcac
     2041 accgtttcta tagattagct aatgagtgtg ctcaagtatt gagtgaaatg gtcatgtgtg
     2101 gcggttcact atatgttaaa ccaggtggaa cctcatcagg agatgccaca actgcttatg
     2161 ctaatagtgt ttttaacatt tgtcaagctg tcacggccaa tgttaatgca cttttatcta
     2221 ctgatggtaa caaaattgcc gataagtatg tccgcaattt acaacacaga ctttatgagt
     2281 gtctctatag aaatagagat gttgacacag actttgtgaa tgagttttac gcatatttgc
     2341 gtaaacattt ctcaatgatg atactctctg acgatgctgt tgtgtgtttc aatagcactt
     2401 atgcatctca aggtctagtg gctagcataa agaactttaa gtcagttctt tattatcaaa
     2461 acaatgtttt tatgtctgaa gcaaaatgtt ggactgagac tgaccttact aaaggacctc
     2521 atgaattttg ctctcaacat acaatgctag ttaaacaggg tgatgattat gtgtaccttc
     2581 cttacccaga tccatcaaga atcctagggg ccggctgttt tgtagatgat atcgtaaaaa
     2641 cagatggtac acttatgatt gaacggttcg tgtctttagc tatagatgct tacccactta
     2701 ctaaacatcc taatcaggag tatgctgatg tctttcattt gtacttacaa tacataagaa
     2761 agctacatga tgagttaaca ggacacatgt tagacatgta ttctgttatg cttactaatg
     2821 ataacacttc aaggtattgg gaacctgagt tttatgaggc tatgtacaca ccgcatacag
     2881 tcttagagaa cctgtacttt caaagcgagc agaagcttat ttccgaagaa gatctctaac
     2941 cggcttatcg gtcagtttca cctgatttac gtaaaaaccc gcttcggcgg gtttttgctt
     3001 ttggaggggc agaaagatga atgactgtcc acgacgctat acccaaaaga aaaagcttga
     3061 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3121 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3181 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3241 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3301 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3361 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3421 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3481 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3541 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3601 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaattgat cgggcacgta
     3661 agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta
     3721 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
     3781 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
     3841 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
     3901 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
     3961 ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
     4021 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
     4081 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
     4141 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtttcagc caatccctgg
     4201 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     4261 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     4321 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     4381 tactgcgatg agtggcaggg cggggcgtaa tttgatatcg agctcgcttg gactcctgtt
     4441 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg
     4501 ccgggcgttt tttattggtc atcgatctcg atcccgcg
//
