LOCUS       sequence-295578-        8150 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 8150)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8150)
  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-m4134228
FEATURES             Location/Qualifiers
     source          1..8150
                     /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    1690..1811
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1701..1720)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1755..1774
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1926..1972
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2002..2138
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2183..2254
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2480..3068)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2560..2579)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3568..4777
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4614..4633
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4781..5809
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5836..5884
                     /note="synthetic polyadenylation signal"
     CDS             join(6150..6218,6219..7010)
                     /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(6368..6387)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7023..7159
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7304..7375
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7380..7426)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7444..7492
                     /note="synthetic polyadenylation signal"
     enhancer        7573..7876
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7878..8080
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8031..8051
                     /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 ataaggatcc gcaggcctct gctagcttga ctgactgaga tacagcgtac cttcagctca
     1681 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     1741 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     1801 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     1861 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg
     1921 tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc
     1981 gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc
     2041 gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac
     2101 ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc
     2161 gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt
     2221 cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg
     2281 aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact
     2341 acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga
     2401 ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2461 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2521 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2581 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2641 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2701 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2761 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2821 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2881 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2941 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3001 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3061 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3121 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3181 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3241 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3301 ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc
     3361 cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc
     3421 gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga
     3481 gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc
     3541 gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg
     3601 gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga
     3661 tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc
     3721 ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt
     3781 ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg
     3841 atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct
     3901 agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca
     3961 cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt
     4021 gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg
     4081 ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg
     4141 gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg
     4201 gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat
     4261 gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt
     4321 tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt
     4381 tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg
     4441 gtggggccac ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc
     4501 gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata
     4561 agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct
     4621 ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc
     4681 ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc
     4741 cgctaaattc tggccgtttt tggctttttt gttagacaac atgggtaaaa agcctgaact
     4801 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
     4861 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
     4921 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
     4981 ctttgcatcc gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag
     5041 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
     5101 cgaactgccc gctgttctgc agccggtcgc ggaggcaatg gatgccatcg ctgccgccga
     5161 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     5221 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     5281 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     5341 ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac
     5401 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     5461 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
     5521 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccccggctcc gggcgtatat
     5581 gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc
     5641 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
     5701 tacacaaatc gcccgcagaa gcgccgccgt ctggaccgat ggctgtgtag aagtactcgc
     5761 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaataag ctagtatgta
     5821 agcctagtct tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg
     5881 tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     5941 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     6001 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     6061 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     6121 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     6181 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6241 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     6301 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     6361 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6421 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     6481 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6541 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     6601 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6661 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     6721 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     6781 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6841 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6901 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6961 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     7021 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     7081 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     7141 accagaaatt tatccttaag atcgtttaaa ctcgactctg gctctatcga atctccgtcg
     7201 tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca
     7261 gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc
     7321 tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc
     7381 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt
     7441 acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact
     7501 gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata
     7561 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     7621 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     7681 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     7741 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     7801 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     7861 catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     7921 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     7981 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     8041 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     8101 cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
