LOCUS       sequence-295147-        7985 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 7985)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7985)
  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-m4133190
FEATURES             Location/Qualifiers
     source          1..7985
                     /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             1415..1432
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1525..1646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1536..1555)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1590..1609
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1761..1807
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1837..1973
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2018..2089
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2315..2903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2395..2414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3403..4612
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4449..4468
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4616..5644
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5671..5719
                     /note="synthetic polyadenylation signal"
     CDS             join(5985..6053,6054..6845)
                     /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(6203..6222)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6858..6994
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7139..7210
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7215..7261)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7279..7327
                     /note="synthetic polyadenylation signal"
     enhancer        7408..7711
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7713..7915
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7866..7886
                     /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 cagtcgatga gtgacaaaat
     1081 cattcatcta accgatgact cctttgatac agacgtgttg aaggccgatg gtgcgatact
     1141 ggtagatttc tgggctgaat ggtgtggccc gtgcaagatg atcgcaccca ttctggacga
     1201 gatcgccgac gagtatcaag gaaaattaac tgtcgctaag cttaatatag atcagaaccc
     1261 agggacggca cctaaatacg gcatcagggg aatacctacc ctcctgctct ttaagaatgg
     1321 ggaagttgcc gcgacaaaag tgggagccct gtctaagggc cagctaaaag agttcttgga
     1381 cgctaacctg gcaggttcag gctcggggca catgcaccat caccatcacc atagcagcgg
     1441 gctcgtgcca cgcggctaag gatccgcagg cctctgctag cttgactgac tgagatacag
     1501 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
     1561 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     1621 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     1681 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
     1741 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1801 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
     1861 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
     1921 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
     1981 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
     2041 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
     2101 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
     2161 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
     2221 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
     2281 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2341 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2401 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2461 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2521 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2581 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2641 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2701 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2761 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2821 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2881 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2941 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3001 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3061 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3121 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     3181 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     3241 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     3301 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     3361 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt
     3421 tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc
     3481 gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     3541 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     3601 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     3661 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     3721 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     3781 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     3841 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     3901 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     3961 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     4021 ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc
     4081 ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga
     4141 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca
     4201 gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt
     4261 ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg
     4321 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg
     4381 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct
     4441 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga
     4501 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta
     4561 gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatggg
     4621 taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     4681 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     4741 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     4801 ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc ttgacattgg
     4861 ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     4921 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg caatggatgc
     4981 catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     5041 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     5101 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     5161 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     5221 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     5281 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     5341 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccccg
     5401 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     5461 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     5521 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga ccgatggctg
     5581 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     5641 ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg
     5701 tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct
     5761 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     5821 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     5881 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     5941 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     6001 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6061 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     6121 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     6181 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6241 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     6301 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6361 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     6421 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6481 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6541 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     6601 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6661 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6721 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6781 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6841 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     6901 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     6961 atacgtgtcc tccttaccag aaatttatcc ttaagatcgt ttaaactcga ctctggctct
     7021 atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg
     7081 catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca
     7141 tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca
     7201 gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
     7261 atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga
     7321 ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt
     7381 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     7441 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     7501 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     7561 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     7621 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     7681 catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg
     7741 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     7801 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     7861 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     7921 agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg
     7981 gccgc
//
