LOCUS       sequence-296392-        4532 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 4532)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4532)
  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-m4046940
FEATURES             Location/Qualifiers
     source          1..4532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /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             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2863..2883
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2884..2907
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      3028..3616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3517..3536
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3722..4381
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3788..3807)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4452..4486
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     primer_bind     join(4514..4532,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4514..4532
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gactcttcta tggctgatgc acaatcgttt ttaaaccggg tttgcggtgt aagtgcagcc
      121 cgtcttacac cgtgcggcac aggcactagt actgatgtcg tatacagggc ttttgacatc
      181 tacaatgata aagtagctgg ttttgctaaa ttcctaaaaa ctaattgttg tcgcttccaa
      241 gaaaaggacg aagatgacaa tttaattgat tcttactttg tagttaagag acacactttc
      301 tctaactacc aacatgaaga aacaatttat aatttactta aggattgtcc agctgttgct
      361 aaacatgact tctttaagtt tagaatagac ggtgacatgg taccacatat atcacgtcaa
      421 cgtcttacta aatacacaat ggcagacctc gtctatgctt taaggcattt tgatgaaggt
      481 aattgtgaca cattaaaaga aatacttgtc acatacaatt gttgtgatga tgattatttc
      541 aataaaaagg actggtatga ttttgtagaa aacccagata tattacgcgt atacgccaac
      601 ttaggtgaac gtgtacgcca agctttgtta aaaacagtac aattctgtga tgccatgcga
      661 aatgctggta ttgttggtgt actgacatta gataatcaag atctcaatgg taactggtat
      721 gatttcggtg atttcataca aaccacgcca ggtagtggag ttcctgttgt agattcttat
      781 tattcattgt taatgcctat attaaccttg accagggctt taactgcaga gtcacatgtt
      841 gacactgact taacaaagcc ttacattaag tgggatttgt taaaatatga cttcacggaa
      901 gagaggttaa aactctttga ccgttatttt aaatattggg atcagacata ccacccaaat
      961 tgtgttaact gtttggatga cagatgcatt ctgcattgtg caaactttaa tgttttattc
     1021 tctacagtgt tcccacctac aagttttgga ccactagtga gaaaaatatt tgttgatggt
     1081 gttccatttg tagtttcaac tggataccac ttcagagagc taggtgttgt acataatcag
     1141 gatgtaaact tacatagctc tagacttagt tttaaggaat tacttgtgta tgctgctgac
     1201 cctgctatgc acgctgcttc tggtaatcta ttactagata aacgcactac gtgcttttca
     1261 gtagctgcac ttactaacaa tgttgctttt caaactgtca aacccggtaa ttttaacaaa
     1321 gacttctatg actttgctgt gtctaagggt ttctttaagg aaggaagttc tgttgaatta
     1381 aaacacttct tctttgctca ggatggtaat gctgctatca gcgattatga ctactatcgt
     1441 tataatctac caacaatgtg tgatatcaga caactactat ttgtagttga agttgttgat
     1501 aagtactttg attgttacga tggtggctgt attaatgcta accaagtcat cgtcaacaac
     1561 ctagacaaat cagctggttt tccatttaat aaatggggta aggctagact ttattatgat
     1621 tcaatgagtt atgaggatca agatgcactt ttcgcatata caaaacgtaa tgtcatccct
     1681 actataactc aaatgaatct taagtatgcc attagtgcaa agaatagagc tcgcaccgta
     1741 gctggtgtct ctatctgtag tactatgacc aatagacagt ttcatcaaaa attattgaaa
     1801 tcaatagccg ccactagagg agctactgta gtaattggaa caagcaaatt ctatggtggt
     1861 tggcacaaca tgttaaaaac tgtttatagt gatgtagaaa accctcacct tatgggttgg
     1921 gattatccta aatgtgatag agccatgcct aacatgctta gaattatggc ctcacttgtt
     1981 cttgctcgca aacatacaac gtgttgtagc ttgtcacacc gtttctatag attagctaat
     2041 gagtgtgctc aagtattgag tgaaatggtc atgtgtggcg gttcactata tgttaaacca
     2101 ggtggaacct catcaggaga tgccacaact gcttatgcta atagtgtttt taacatttgt
     2161 caagctgtca cggccaatgt taatgcactt ttatctactg atggtaacaa aattgccgat
     2221 aagtatgtcc gcaatttaca acacagactt tatgagtgtc tctatagaaa tagagatgtt
     2281 gacacagact ttgtgaatga gttttacgca tatttgcgta aacatttctc aatgatgata
     2341 ctctctgacg atgctgttgt gtgtttcaat agcacttatg catctcaagg tctagtggct
     2401 agcataaaga actttaagtc agttctttat tatcaaaaca atgtttttat gtctgaagca
     2461 aaatgttgga ctgagactga ccttactaaa ggacctcatg aattttgctc tcaacataca
     2521 atgctagtta aacagggtga tgattatgtg taccttcctt acccagatcc atcaagaatc
     2581 ctaggggccg gctgttttgt agatgatatc gtaaaaacag atggtacact tatgattgaa
     2641 cggttcgtgt ctttagctat agatgcttac ccacttacta aacatcctaa tcaggagtat
     2701 gctgatgtct ttcatttgta cttacaatac ataagaaagc tacatgatga gttaacagga
     2761 cacatgttag acatgtattc tgttatgctt actaatgata acacttcaag gtattgggaa
     2821 cctgagtttt atgaggctat gtacacaccg catacagtct tagagaacct gtactttcaa
     2881 agctggagcc atccgcagtt tgaaaaataa ccggcttatc ggtcagtttc acctgattta
     2941 cgtaaaaacc cgcttcggcg ggtttttgct tttggagggg cagaaagatg aatgactgtc
     3001 cacgacgcta tacccaaaag aaaaagcttg agatcctttt tttctgcgcg taatctgctg
     3061 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3121 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3181 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3241 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3301 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3361 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3421 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3481 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3541 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3601 gcggagccta tggaaattga tcgggcacgt aagaggttcc aactttcacc ataatgaaat
     3661 aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa
     3721 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     3781 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     3841 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     3901 tcacattctt gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg
     3961 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     4021 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     4081 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     4141 gaatatgttt ttcgtttcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     4201 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     4261 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     4321 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     4381 atttgatatc gagctcgctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
     4441 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt catcgatctc
     4501 gatcccgcga aattaatacg actcactata gg
//
