LOCUS       sequence-295407-        8144 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 8144)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8144)
  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-m4133133
FEATURES             Location/Qualifiers
     source          1..8144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1040..1060
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1094..1171
                     /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             1199..1219
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1684..1805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1695..1714)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1749..1768
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1920..1966
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1996..2132
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2177..2248
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2474..3062)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2554..2573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3562..4771
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4608..4627
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4775..5803
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5830..5878
                     /note="synthetic polyadenylation signal"
     CDS             join(6144..6212,6213..7004)
                     /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(6362..6381)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7017..7153
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7298..7369
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7374..7420)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7438..7486
                     /note="synthetic polyadenylation signal"
     enhancer        7567..7870
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7872..8074
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8025..8045
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtta gaaaatgtgg cttttaatgt tgtaaataag ggacactttg atggacaaca
       61 gggtgaagta ccagtttcta tcattaataa cactgtttac acaaaagttg atggtgttga
      121 tgtagaattg tttgaaaata aaacaacatt acctgttaat gtagcatttg agctttgggc
      181 taagcgcaac attaaaccag taccagaggt gaaaatactc aataatttgg gtgtggacat
      241 tgctgctaat actgtgatct gggactacaa aagagatgct ccagcacata tatctactat
      301 tggtgtttgt tctatgactg acatagccaa gaaaccaact gaaacgattt gtgcaccact
      361 cactgtcttt tttgatggta gagttgatgg tcaagtagac ttatttagaa atgcccgtaa
      421 tggtgttctt attacagaag gtagtgttaa aggtttacaa ccatctgtag gtcccaaaca
      481 agctagtctt aatggagtca cattaattgg agaagccgta aaaacacagt tcaattatta
      541 taagaaagtt gatggtgttg tccaacaatt acctgaaact tactttactc agagtagaaa
      601 tttacaagaa tttaaaccca ggagtcaaat ggaaattgat ttcttagaat tagctatgga
      661 tgaattcatt gaacggtata aattagaagg ctatgccttc gaacatatcg tttatggaga
      721 ttttagtcat agtcagttag gtggtttaca tctactgatt ggactagcta aacgttttaa
      781 ggaatcacct tttgaattag aagattttat tcctatggac agtacagtta aaaactattt
      841 cataacagat gcgcaaacag gttcatctaa gtgtgtgtgt tctgttattg atttattact
      901 tgatgatttt gttgaaataa taaaatccca agatttatct gtagtttcta aggttgtcaa
      961 agtgactatt gactatacag aaatttcatt tatgctttgg tgtaaagatg gccatgtaga
     1021 aacattttac ccaaaattag aaaaccttta ctttcagtcg ggacggcgaa tcccggggct
     1081 gataaatccc tggaaaagga gatggaagaa gaactttatt gcagtgtctg cggctaaccg
     1141 gttcaaaaag atcagttcaa gcggcgccct agattacgac attcctacga ccgcctccga
     1201 gaatctctat ttccagggtg aatttggctt ggcacaacat gacgaggccg tcgataacaa
     1261 gtttaataaa gaacagcaga acgctttcta tgagatcctt cacttaccaa atctgaacga
     1321 agagcaacgc aacgctttca ttcagagcct gaaagacgat ccctcgcagt ctgcaaatct
     1381 cctggccgaa gccaagaagc tgaatgacgc acaggctcca aaagttgata acaagtttaa
     1441 caaggagcag cagaatgctt tctacgagat attgcacctt cctaatctga atgaggaaca
     1501 aagaaatgca tttatccagt ccctaaaaga cgatccatca cagagtgcca acctgcttgc
     1561 ggaagccaaa aagctcaatg atgcccaggc cccgaaagta gacgcgaacc atcaataagg
     1621 atccgcaggc ctctgctagc ttgactgact gagatacagc gtaccttcag ctcacagaca
     1681 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     1741 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     1801 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     1861 ttttttaaag caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt
     1921 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg
     1981 gattgctatc taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta
     2041 tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat
     2101 acgtgtcctc cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc
     2161 tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag
     2221 tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt
     2281 aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc
     2341 agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga
     2401 gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2461 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2521 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2581 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2641 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2701 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2761 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2821 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2881 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2941 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3001 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3061 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3121 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3181 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3241 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat
     3301 ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc
     3361 atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg
     3421 gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg
     3481 gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa
     3541 tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc
     3601 ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc
     3661 cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta
     3721 tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc
     3781 agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc
     3841 gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc
     3901 gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt
     3961 gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg
     4021 gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga
     4081 actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac
     4141 gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga
     4201 acccaaggtc ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg
     4261 ggcaccatct ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt
     4321 ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag
     4381 cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg
     4441 ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct
     4501 agggtaggct ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag
     4561 gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt
     4621 ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga
     4681 aatgtaatca tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa
     4741 attctggccg tttttggctt ttttgttaga caacatgggt aaaaagcctg aactcaccgc
     4801 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
     4861 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
     4921 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
     4981 atccgccgcg ctcccgattc cggaagtgct tgacattggg gagttcagcg agagcctgac
     5041 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
     5101 gcccgctgtt ctgcagccgg tcgcggaggc aatggatgcc atcgctgccg ccgatcttag
     5161 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
     5221 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
     5281 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
     5341 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
     5401 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
     5461 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
     5521 cttcgagcgg aggcatccgg agcttgcagg atcgccccgg ctccgggcgt atatgctccg
     5581 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
     5641 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
     5701 aatcgcccgc agaagcgccg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
     5761 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa taagctagta tgtaagccta
     5821 gtcttagata ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg
     5881 taacgctagg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc
     5941 atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg
     6001 tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt
     6061 gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga
     6121 caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct
     6181 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6241 atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta
     6301 agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc
     6361 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     6421 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg
     6481 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6541 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca
     6601 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6661 acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa
     6721 ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg gaggcggata
     6781 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6841 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6901 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6961 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat
     7021 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     7081 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     7141 aatttatcct taagatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga
     7201 gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc
     7261 gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca
     7321 ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa
     7381 cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac
     7441 acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat
     7501 cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga
     7561 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     7621 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     7681 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     7741 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     7801 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     7861 tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     7921 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     7981 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     8041 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc
     8101 tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
