LOCUS       sequence-307396-        4532 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for 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-m4046955
FEATURES             Location/Qualifiers
     source          1..4532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2748..2768
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2769..2792
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      2913..3501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3402..3421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3607..4266
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3673..3692)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4337..4371
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     primer_bind     4399..4418
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4399..4417
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4418..4442
                     /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             4457..4479
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 cagcccgtct tacaccgtgc ggcacaggca ctagtactga tgtcgtatac agggcttttg
       61 acatctacaa tgataaagta gctggttttg ctaaattcct aaaaactaat tgttgtcgct
      121 tccaagaaaa ggacgaagat gacaatttaa ttgattctta ctttgtagtt aagagacaca
      181 ctttctctaa ctaccaacat gaagaaacaa tttataattt acttaaggat tgtccagctg
      241 ttgctaaaca tgacttcttt aagtttagaa tagacggtga catggtacca catatatcac
      301 gtcaacgtct tactaaatac acaatggcag acctcgtcta tgctttaagg cattttgatg
      361 aaggtaattg tgacacatta aaagaaatac ttgtcacata caattgttgt gatgatgatt
      421 atttcaataa aaaggactgg tatgattttg tagaaaaccc agatatatta cgcgtatacg
      481 ccaacttagg tgaacgtgta cgccaagctt tgttaaaaac agtacaattc tgtgatgcca
      541 tgcgaaatgc tggtattgtt ggtgtactga cattagataa tcaagatctc aatggtaact
      601 ggtatgattt cggtgatttc atacaaacca cgccaggtag tggagttcct gttgtagatt
      661 cttattattc attgttaatg cctatattaa ccttgaccag ggctttaact gcagagtcac
      721 atgttgacac tgacttaaca aagccttaca ttaagtggga tttgttaaaa tatgacttca
      781 cggaagagag gttaaaactc tttgaccgtt attttaaata ttgggatcag acataccacc
      841 caaattgtgt taactgtttg gatgacagat gcattctgca ttgtgcaaac tttaatgttt
      901 tattctctac agtgttccca cctacaagtt ttggaccact agtgagaaaa atatttgttg
      961 atggtgttcc atttgtagtt tcaactggat accacttcag agagctaggt gttgtacata
     1021 atcaggatgt aaacttacat agctctagac ttagttttaa ggaattactt gtgtatgctg
     1081 ctgaccctgc tatgcacgct gcttctggta atctattact agataaacgc actacgtgct
     1141 tttcagtagc tgcacttact aacaatgttg cttttcaaac tgtcaaaccc ggtaatttta
     1201 acaaagactt ctatgacttt gctgtgtcta agggtttctt taaggaagga agttctgttg
     1261 aattaaaaca cttcttcttt gctcaggatg gtaatgctgc tatcagcgat tatgactact
     1321 atcgttataa tctaccaaca atgtgtgata tcagacaact actatttgta gttgaagttg
     1381 ttgataagta ctttgattgt tacgatggtg gctgtattaa tgctaaccaa gtcatcgtca
     1441 acaacctaga caaatcagct ggttttccat ttaataaatg gggtaaggct agactttatt
     1501 atgattcaat gagttatgag gatcaagatg cacttttcgc atatacaaaa cgtaatgtca
     1561 tccctactat aactcaaatg aatcttaagt atgccattag tgcaaagaat agagctcgca
     1621 ccgtagctgg tgtctctatc tgtagtacta tgaccaatag acagtttcat caaaaattat
     1681 tgaaatcaat agccgccact agaggagcta ctgtagtaat tggaacaagc aaattctatg
     1741 gtggttggca caacatgtta aaaactgttt atagtgatgt agaaaaccct caccttatgg
     1801 gttgggatta tcctaaatgt gatagagcca tgcctaacat gcttagaatt atggcctcac
     1861 ttgttcttgc tcgcaaacat acaacgtgtt gtagcttgtc acaccgtttc tatagattag
     1921 ctaatgagtg tgctcaagta ttgagtgaaa tggtcatgtg tggcggttca ctatatgtta
     1981 aaccaggtgg aacctcatca ggagatgcca caactgctta tgctaatagt gtttttaaca
     2041 tttgtcaagc tgtcacggcc aatgttaatg cacttttatc tactgatggt aacaaaattg
     2101 ccgataagta tgtccgcaat ttacaacaca gactttatga gtgtctctat agaaatagag
     2161 atgttgacac agactttgtg aatgagtttt acgcatattt gcgtaaacat ttctcaatga
     2221 tgatactctc tgacgatgct gttgtgtgtt tcaatagcac ttatgcatct caaggtctag
     2281 tggctagcat aaagaacttt aagtcagttc tttattatca aaacaatgtt tttatgtctg
     2341 aagcaaaatg ttggactgag actgacctta ctaaaggacc tcatgaattt tgctctcaac
     2401 atacaatgct agttaaacag ggtgatgatt atgtgtacct tccttaccca gatccatcaa
     2461 gaatcctagg ggccggctgt tttgtagatg atatcgtaaa aacagatggt acacttatga
     2521 ttgaacggtt cgtgtcttta gctatagatg cttacccact tactaaacat cctaatcagg
     2581 agtatgctga tgtctttcat ttgtacttac aatacataag aaagctacat gatgagttaa
     2641 caggacacat gttagacatg tattctgtta tgcttactaa tgataacact tcaaggtatt
     2701 gggaacctga gttttatgag gctatgtaca caccgcatac agtcttagag aacctgtact
     2761 ttcaaagcga ctacaaggat gatgatgaca aataaccggc ttatcggtca gtttcacctg
     2821 atttacgtaa aaacccgctt cggcgggttt ttgcttttgg aggggcagaa agatgaatga
     2881 ctgtccacga cgctataccc aaaagaaaaa gcttgagatc ctttttttct gcgcgtaatc
     2941 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3001 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     3061 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3121 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3181 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3241 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3301 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3361 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3421 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3481 ggggggcgga gcctatggaa attgatcggg cacgtaagag gttccaactt tcaccataat
     3541 gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa
     3601 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     3661 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     3721 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     3781 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa
     3841 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     3901 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     3961 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     4021 attgagaata tgtttttcgt ttcagccaat ccctgggtga gtttcaccag ttttgattta
     4081 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     4141 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     4201 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     4261 gcgtaatttg atatcgagct cgcttggact cctgttgata gatccagtaa tgacctcaga
     4321 actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtcatcg
     4381 atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt
     4441 cccctctaga aataattttg tttaacttta agaaggagac tcttctatgg ctgatgcaca
     4501 atcgttttta aaccgggttt gcggtgtaag tg
//
