LOCUS       sequence-297409-        4278 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4278)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4278)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4133395
FEATURES             Location/Qualifiers
     source          1..4278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             47..67
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             68..94
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    147..268
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(158..177)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     212..231
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      383..429
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      459..595
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    640..711
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(937..1525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1017..1036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2271..2339,2340..3131)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2489..2508)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3144..3280
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3432..3503
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3508..3554)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3572..3620
                     /note="synthetic polyadenylation signal"
     enhancer        3701..4004
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4006..4208
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4159..4179
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt
       61 tcagtcgtac ccctatgacg taccagatta tgcgtaagct agcttgactg actgagatac
      121 agcgtacctt cagctcacag acatgataag atacattgat gagtttggac aaaccacaac
      181 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
      241 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
      301 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
      361 tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg
      421 ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc
      481 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
      541 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg
      601 tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc
      661 tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg
      721 gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac
      781 ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta
      841 ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa
      901 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      961 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1021 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1081 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1141 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1201 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1261 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1321 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1381 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1441 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1501 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1561 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1621 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1681 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1741 atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa
     1801 tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt
     1861 ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc
     1921 caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc
     1981 cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggtgta ccgagaacga
     2041 tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc
     2101 cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc
     2161 gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc
     2221 tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc
     2281 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2341 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt
     2401 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct
     2461 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2521 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt
     2581 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2641 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga
     2701 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2761 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2821 tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2881 agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2941 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3001 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3061 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3121 agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     3181 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     3241 acttccatac gtgtcctcct taccagaaat ttatccttaa gatcccgaat cgtttaaact
     3301 cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca
     3361 tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc
     3421 gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat
     3481 aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc
     3541 aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt
     3601 cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa
     3661 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     3721 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     3781 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     3841 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     3901 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     3961 ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc
     4021 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     4081 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     4141 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     4201 gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac
     4261 acacccgcca gcggccgc
//
