LOCUS       sequence-295588-        7664 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 7664)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7664)
  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-m4134190
FEATURES             Location/Qualifiers
     source          1..7664
                     /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             1070..1135
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1204..1325
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1215..1234)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1269..1288
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1440..1486
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1516..1652
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1697..1768
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1994..2582)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2074..2093)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3082..4291
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4128..4147
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4295..5323
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5350..5398
                     /note="synthetic polyadenylation signal"
     CDS             join(5664..5732,5733..6524)
                     /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(5882..5901)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6537..6673
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6818..6889
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6894..6940)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6958..7006
                     /note="synthetic polyadenylation signal"
     enhancer        7087..7390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7392..7594
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7545..7565
                     /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 cagtcgatgg attataagga
     1081 ccacgatgga gactacaaag atcatgacat tgattacaag gacgacgatg acaaataagg
     1141 atccgcaggc ctctgctagc ttgactgact gagatacagc gtaccttcag ctcacagaca
     1201 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     1261 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     1321 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     1381 ttttttaaag caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt
     1441 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg
     1501 gattgctatc taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta
     1561 tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat
     1621 acgtgtcctc cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc
     1681 tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag
     1741 tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt
     1801 aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc
     1861 agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga
     1921 gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1981 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2041 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2101 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2161 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2221 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2281 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2341 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2401 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2461 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2521 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2581 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2641 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2701 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2761 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat
     2821 ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc
     2881 atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg
     2941 gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg
     3001 gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa
     3061 tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc
     3121 ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc
     3181 cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta
     3241 tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc
     3301 agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc
     3361 gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc
     3421 gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt
     3481 gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg
     3541 gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga
     3601 actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac
     3661 gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga
     3721 acccaaggtc ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg
     3781 ggcaccatct ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt
     3841 ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag
     3901 cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg
     3961 ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct
     4021 agggtaggct ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag
     4081 gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt
     4141 ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga
     4201 aatgtaatca tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa
     4261 attctggccg tttttggctt ttttgttaga caacatgggt aaaaagcctg aactcaccgc
     4321 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
     4381 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
     4441 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
     4501 atccgccgcg ctcccgattc cggaagtgct tgacattggg gagttcagcg agagcctgac
     4561 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
     4621 gcccgctgtt ctgcagccgg tcgcggaggc aatggatgcc atcgctgccg ccgatcttag
     4681 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
     4741 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
     4801 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
     4861 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
     4921 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
     4981 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
     5041 cttcgagcgg aggcatccgg agcttgcagg atcgccccgg ctccgggcgt atatgctccg
     5101 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
     5161 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
     5221 aatcgcccgc agaagcgccg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
     5281 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa taagctagta tgtaagccta
     5341 gtcttagata ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg
     5401 taacgctagg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc
     5461 atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg
     5521 tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt
     5581 gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga
     5641 caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct
     5701 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5761 atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta
     5821 agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc
     5881 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5941 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg
     6001 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6061 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca
     6121 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6181 acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa
     6241 ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg gaggcggata
     6301 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6361 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6421 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6481 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat
     6541 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     6601 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     6661 aatttatcct taagatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga
     6721 gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc
     6781 gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca
     6841 ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa
     6901 cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac
     6961 acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat
     7021 cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga
     7081 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     7141 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     7201 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     7261 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     7321 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     7381 tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     7441 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     7501 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     7561 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc
     7621 tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
