LOCUS       sequence-295908-        7634 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 7634)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7634)
  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-m4133432
FEATURES             Location/Qualifiers
     source          1..7634
                     /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             1061..1105
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1174..1295
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1185..1204)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1239..1258
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1410..1456
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1486..1622
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1667..1738
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1964..2552)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2044..2063)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3052..4261
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4098..4117
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4265..5293
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5320..5368
                     /note="synthetic polyadenylation signal"
     CDS             join(5634..5702,5703..6494)
                     /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(5852..5871)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6507..6643
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6788..6859
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6864..6910)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6928..6976
                     /note="synthetic polyadenylation signal"
     enhancer        7057..7360
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7362..7564
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7515..7535
                     /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 aaagaaaccg cggcagctaa
     1081 gttcgagagg cagcacatgg acagttaagg atccgcaggc ctctgctagc ttgactgact
     1141 gagatacagc gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa
     1201 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     1261 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1321 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     1381 gtggtattgg cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt
     1441 cttgaggggt tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa
     1501 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
     1561 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
     1621 aaggtcgtca gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg
     1681 tcttttctgg acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat
     1741 ctcattggtt accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga
     1801 gatcgacttg tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc
     1861 cttgctattg cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc
     1921 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1981 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2041 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2101 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2161 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2221 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2281 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2341 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2401 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2461 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2521 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2581 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2641 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2701 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2761 tctgtctatt tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt
     2821 cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat
     2881 cgcaagtctc ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt
     2941 attactccaa tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc
     3001 tcaatcccga tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc
     3061 gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag
     3121 acgaagggcg caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg
     3181 ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt
     3241 gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc
     3301 ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa
     3361 ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt
     3421 gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct
     3481 ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt
     3541 agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg
     3601 gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa
     3661 acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg
     3721 ggaaagctct tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt
     3781 tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc
     3841 cgttgggcag tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc
     3901 cgttctgttg gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt
     3961 tctccgtcgc aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc
     4021 ggacctctgg tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac
     4081 ctatcttctt aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt
     4141 gttttgtgaa gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt
     4201 tcagtgttag acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga
     4261 caacatgggt aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     4321 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     4381 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     4441 caaagatcgt tatgtttatc ggcactttgc atccgccgcg ctcccgattc cggaagtgct
     4501 tgacattggg gagttcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     4561 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     4621 aatggatgcc atcgctgccg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     4681 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     4741 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     4801 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     4861 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     4921 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     4981 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     5041 atcgccccgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     5101 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     5161 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgccg ccgtctggac
     5221 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     5281 ggcaaaggaa taagctagta tgtaagccta gtcttagata ataaaatcgc tatccatcga
     5341 agatggatgt gtgttggttt tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa
     5401 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     5461 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     5521 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     5581 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     5641 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5701 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     5761 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5821 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5881 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5941 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     6001 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     6061 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     6121 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     6181 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     6241 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6301 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6361 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6421 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6481 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     6541 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     6601 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcgtt taaactcgac
     6661 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     6721 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     6781 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     6841 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     6901 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     6961 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     7021 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     7081 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     7141 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     7201 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     7261 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     7321 ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt tttggcagta
     7381 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     7441 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     7501 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     7561 agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca ctcgacacac
     7621 ccgccagcgg ccgc
//
