LOCUS       sequence-295421-        5805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 endoRNAse.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5805)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5805)
  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-m4133194
FEATURES             Location/Qualifiers
     source          1..5805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1046..1066
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1100..1177
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             1205..1225
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1674..1795
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1685..1704)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1739..1758
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1910..1956
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1986..2122
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2167..2238
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2464..3052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2544..2563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3798..3866,3867..4658)
                     /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(4016..4035)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4671..4807
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4959..5030
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5035..5081)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5099..5147
                     /note="synthetic polyadenylation signal"
     enhancer        5228..5531
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5533..5735
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5686..5706
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtca ctggagaacg ttgctttcaa tgtagtgaac aaggggcact ttgacggaca
       61 gcaaggcgaa gtgccggtca gtatcataaa taatacggtg tacaccaaag tcgatggtgt
      121 tgacgtggag ttatttgaaa acaagacaac tttgcctgtc aatgtggcct tcgagctgtg
      181 ggcaaaacgc aacattaagc cagtacccga agtgaaaatc ctcaacaatc ttggcgttga
      241 tattgccgca aacactgtta tctgggatta taagcgagat gcgcctgccc atatatccac
      301 cattggagtg tgttctatga cagacatcgc taaaaagccc accgagacaa tttgcgcgcc
      361 actcacggta ttctttgacg ggagggtcga cggtcaggtg gatctgttta gaaatgcccg
      421 taacggcgta cttattactg aaggaagcgt gaaaggccta cagcccagtg ttgggccgaa
      481 gcaagcaagc ctaaatggag tcacactgat cggagaggct gtgaaaaccc agttcaacta
      541 ctataagaag gtcgacggcg tggttcagca gctgcctgag acatacttta cccaatcgcg
      601 gaatctgcaa gaatttaagc ccagatccca gatggagatt gacttcttgg aactggccat
      661 ggatgagttc atcgaacggt ataagctcga gggctacgcc ttcgagcaca tagtttatgg
      721 ggatttttct cattcacagt taggcggtct gcacttgctc atcggattag cgaaacgctt
      781 taaagagagc ccattcgaac ttgaagactt tatcccaatg gactcaactg tgaagaatta
      841 cttcataact gacgctcaga cagggtccag caagtgcgtc tgttctgtta ttgatttgct
      901 acttgatgat ttcgtggaga ttattaaatc acaggacctg tccgtggttt cgaaggtcgt
      961 gaaagtaacc atcgactata cggaaattag tttcatgctt tggtgtaagg atggccatgt
     1021 ggagacattt tatcctaagc tccaggaaaa cctttacttt cagtcgggac ggcgaatccc
     1081 ggggctgata aatccctgga aaaggagatg gaagaagaac tttattgcag tgtctgcggc
     1141 taaccggttc aaaaagatca gttcaagcgg cgccctagat tacgacattc ctacgaccgc
     1201 ctccgagaat ctctatttcc agggtgaatt tggcttggca caacatgacg aggccgtcga
     1261 taacaagttt aataaagaac agcagaacgc tttctatgag atccttcact taccaaatct
     1321 gaacgaagag caacgcaacg ctttcattca gagcctgaaa gacgatccct cgcagtctgc
     1381 aaatctcctg gccgaagcca agaagctgaa tgacgcacag gctccaaaag ttgataacaa
     1441 gtttaacaag gagcagcaga atgctttcta cgagatattg caccttccta atctgaatga
     1501 ggaacaaaga aatgcattta tccagtccct aaaagacgat ccatcacaga gtgccaacct
     1561 gcttgcggaa gccaaaaagc tcaatgatgc ccaggccccg aaagtagacg cgaaccatca
     1621 ataagctagc ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata
     1681 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     1741 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     1801 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     1861 caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc
     1921 cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc
     1981 taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     2041 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     2101 cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga
     2161 tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca
     2221 aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag
     2281 tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt
     2341 cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta
     2401 aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2461 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2521 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2581 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2641 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2701 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2761 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2821 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2881 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2941 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3001 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3061 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3121 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3181 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3241 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc
     3301 gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg
     3361 ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt
     3421 ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga
     3481 gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc
     3541 ggggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     3601 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     3661 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     3721 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     3781 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3841 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3901 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3961 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     4021 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4081 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     4141 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4201 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4261 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4321 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     4381 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     4441 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4501 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4561 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4621 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     4681 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     4741 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     4801 tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg
     4861 agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat
     4921 cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac
     4981 aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa
     5041 acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca
     5101 cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc taactgctga
     5161 tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg
     5221 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     5281 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     5341 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     5401 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     5461 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     5521 ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     5581 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     5641 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     5701 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat
     5761 ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgc
//
