LOCUS       Exported                2630 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 2'-O-ribose 
            methyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046795
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2630)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2630)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2630
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             1005..1025
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1026..1043
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      1164..1752
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1653..1672
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1858..2517
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1924..1943)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2588..2622
                     /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 tcaagccaag
      121 catggcagcc cggggtggcc atgccaaact tgtacaaaat gcagcgtatg ttactggaga
      181 agtgcgacct gcaaaattat ggtgattctg cgacgcttcc taaaggaatt atgatgaacg
      241 tagctaaata cactcagctg tgtcagtatc tcaacaccct gacactggct gtcccgtaca
      301 acatgcgcgt tatccacttc ggcgcaggct ccgataaagg tgttgcgccg ggcactgctg
      361 tactgcgtca gtggctgccg accggtactc tgctggttga ctctgacctc aatgattttg
      421 tgagtgacgc cgattccacc ctgatcggtg attgcgctac cgttcatact gcgaacaaat
      481 gggacctgat tatcagcgat atgtacgacc cgaagaccaa aaacgtaacc aaagaaaacg
      541 actcgaagga aggcttcttt acttatatct gtggtttcat tcagcagaaa ctggctcttg
      601 gcggttcggt ggcaatcaag attacggaac actcttggaa cgctgaccta tacaaactta
      661 tgggccattt cgcgtggtgg accgcctttg ttactaatgt caacgcgagc agctctgagg
      721 cattcctgat cggatgcaat tacctgggta aaccgcgaga acagatcgat ggctatgtta
      781 tgcacgctaa ctacatattc tggcgtaaca cgaacccgat ccaattatcc tcatattccc
      841 tgtttgacat gagcaaattc ccactaaaac tgcgcggtac agcagtgatg tctctgaaag
      901 aaggccagat taacgacatg attctgagtc tgctgtctaa agggagactg atcatccgtg
      961 agaataaccg tgtggtcatc tcatccgatg ttctggtaaa caatgagaac ctgtactttc
     1021 aaagccacca tcatcaccat cattaaccgg cttatcggtc agtttcacct gatttacgta
     1081 aaaacccgct tcggcgggtt tttgcttttg gaggggcaga aagatgaatg actgtccacg
     1141 acgctatacc caaaagaaaa agcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1201 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1261 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     1321 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1381 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1441 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1501 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1561 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1621 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     1681 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     1741 agcctatgga aattgatcgg gcacgtaaga ggttccaact ttcaccataa tgaaataaga
     1801 tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg
     1861 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
     1921 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
     1981 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
     2041 attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag
     2101 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
     2161 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
     2221 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
     2281 atgtttttcg tttcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
     2341 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
     2401 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     2461 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt
     2521 gatatcgagc tcgcttggac tcctgttgat agatccagta atgacctcag aactccatct
     2581 ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtcatc
//
