LOCUS       sequence-295458-        7613 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 7613)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7613)
  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-m4133214
FEATURES             Location/Qualifiers
     source          1..7613
                     /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             1067..1084
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1153..1274
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1164..1183)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1218..1237
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1389..1435
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1465..1601
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1646..1717
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1943..2531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2023..2042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3031..4240
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4077..4096
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4244..5272
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5299..5347
                     /note="synthetic polyadenylation signal"
     CDS             join(5613..5681,5682..6473)
                     /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(5831..5850)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6486..6622
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6767..6838
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6843..6889)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6907..6955
                     /note="synthetic polyadenylation signal"
     enhancer        7036..7339
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7341..7543
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7494..7514
                     /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 cagtcgcacc atcaccatca
     1081 ccattaagga tccgcaggcc tctgctagct tgactgactg agatacagcg taccttcagc
     1141 tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     1201 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     1261 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     1321 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat
     1381 cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc
     1441 ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg
     1501 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
     1561 cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca
     1621 ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg
     1681 ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta
     1741 tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg
     1801 actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc
     1861 cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1921 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1981 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2041 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2101 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2161 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2221 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2281 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2341 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2401 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2461 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2521 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2581 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2641 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2701 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2761 tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc
     2821 gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct
     2881 tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc
     2941 cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga
     3001 tccgaggaat atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg gcgcctcccg
     3061 cgggcgcccc cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc aggagcgtcc
     3121 tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga
     3181 accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt
     3241 tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga
     3301 gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca
     3361 gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg
     3421 tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc
     3481 ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg
     3541 ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc cgagtcttga
     3601 atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg
     3661 gcggcaagaa cccaaggtct tgagcccttc gctaatgcgg gaaagctctt attcgggtga
     3721 gatgggctgg gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg
     3781 gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac
     3841 ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc
     3901 agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg
     3961 ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg
     4021 ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa
     4081 gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc
     4141 accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga cttgtaaatt
     4201 gtccgctaaa ttctggccgt ttttggcttt tttgttagac aacatgggta aaaagcctga
     4261 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     4321 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     4381 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     4441 gcactttgca tccgccgcgc tcccgattcc ggaagtgctt gacattgggg agttcagcga
     4501 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     4561 aaccgaactg cccgctgttc tgcagccggt cgcggaggca atggatgcca tcgctgccgc
     4621 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     4681 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     4741 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     4801 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     4861 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     4921 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     4981 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccccggc tccgggcgta
     5041 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     5101 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     5161 gcgtacacaa atcgcccgca gaagcgccgc cgtctggacc gatggctgtg tagaagtact
     5221 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat aagctagtat
     5281 gtaagcctag tcttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt
     5341 ttgtgtgtgt aacgctaggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     5401 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     5461 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     5521 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     5581 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     5641 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5701 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     5761 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     5821 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5881 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     5941 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6001 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     6061 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6121 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     6181 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     6241 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6301 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6361 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6421 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     6481 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     6541 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     6601 cttaccagaa atttatcctt aagatcgttt aaactcgact ctggctctat cgaatctccg
     6661 tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg
     6721 tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt
     6781 agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa
     6841 ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc
     6901 gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta
     6961 actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc
     7021 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     7081 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     7141 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     7201 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     7261 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     7321 agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     7381 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     7441 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     7501 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
     7561 gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgc
//
