LOCUS       sequence-300757-        4635 bp    DNA     circular SYN 15-DEC-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 4635)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4635)
  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-m4134204
FEATURES             Location/Qualifiers
     source          1..4635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             41..61
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             410..427
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    504..625
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(515..534)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     569..588
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      740..786
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      816..952
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    997..1068
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1294..1882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1374..1393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2628..2696,2697..3488)
                     /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(2846..2865)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3501..3637
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3789..3860
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3865..3911)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3929..3977
                     /note="synthetic polyadenylation signal"
     enhancer        4058..4361
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4363..4565
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4516..4536
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4634..4635,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggct gatgcacaat cgtttttaaa cgggtttgcg gaaaaccttt actttcagtc
       61 gatgagtgac aaaatcattc atctaaccga tgactccttt gatacagacg tgttgaaggc
      121 cgatggtgcg atactggtag atttctgggc tgaatggtgt ggcccgtgca agatgatcgc
      181 acccattctg gacgagatcg ccgacgagta tcaaggaaaa ttaactgtcg ctaagcttaa
      241 tatagatcag aacccaggga cggcacctaa atacggcatc aggggaatac ctaccctcct
      301 gctctttaag aatggggaag ttgccgcgac aaaagtggga gccctgtcta agggccagct
      361 aaaagagttc ttggacgcta acctggcagg ttcaggctcg gggcacatgc accatcacca
      421 tcaccatagc agcgggctcg tgccacgcgg ctaagctagc ttgactgact gagatacagc
      481 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
      541 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
      601 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
      661 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
      721 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
      781 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
      841 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
      901 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
      961 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     1021 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     1081 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     1141 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     1201 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     1261 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1321 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1381 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1441 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     1501 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     1561 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1621 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     1681 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     1741 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     1801 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     1861 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1921 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     1981 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2041 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2101 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     2161 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     2221 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     2281 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     2341 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc
     2401 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     2461 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     2521 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     2581 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     2641 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2701 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     2761 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     2821 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     2881 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     2941 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3001 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     3061 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3121 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3181 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     3241 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3301 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3361 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3421 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3481 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     3541 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     3601 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga
     3661 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     3721 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     3781 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     3841 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     3901 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga
     3961 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta
     4021 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     4081 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     4141 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     4201 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     4261 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     4321 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt
     4381 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     4441 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     4501 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     4561 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca
     4621 cccgccagcg gccgc
//
