LOCUS       sequence-295524-        7622 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 7622)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7622)
  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-m4134206
FEATURES             Location/Qualifiers
     source          1..7622
                     /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..1093
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1162..1283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1173..1192)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1227..1246
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1398..1444
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1474..1610
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1655..1726
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1952..2540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2032..2051)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3040..4249
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4086..4105
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4253..5281
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5308..5356
                     /note="synthetic polyadenylation signal"
     CDS             join(5622..5690,5691..6482)
                     /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(5840..5859)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6495..6631
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6776..6847
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6852..6898)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6916..6964
                     /note="synthetic polyadenylation signal"
     enhancer        7045..7348
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7350..7552
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7503..7523
                     /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 cagtcgtacc cctatgacgt
     1081 accagattat gcgtaaggat ccgcaggcct ctgctagctt gactgactga gatacagcgt
     1141 accttcagct cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     1201 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1261 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1321 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc
     1381 catctctatc ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt
     1441 tttgtgcccc tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat
     1501 gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc
     1561 caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc
     1621 tatcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac
     1681 accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac
     1741 cttgggctat cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc
     1801 tgggtttcga ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca
     1861 cccgttctcc gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca
     1921 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1981 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2041 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2101 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2161 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2221 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2281 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2341 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2401 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2461 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2521 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2581 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2641 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2701 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2761 gttcatccat agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc
     2821 aaacctttcg tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt
     2881 ggccggcctt gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc
     2941 ccgaatatcc gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc
     3001 tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg
     3061 cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca
     3121 ggagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc
     3181 ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag
     3241 ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga
     3301 ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg
     3361 tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg
     3421 ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc
     3481 gggccgctcg gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg
     3541 cgagcaaggt tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc
     3601 gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg
     3661 gcatggtggg cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg aaagctctta
     3721 ttcgggtgag atgggctggg caccatctgg ggaccctgac gtgaagtttg tcactgactg
     3781 gagaactcgg tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg
     3841 cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc
     3901 ttataatgca gggtggggcc acctgccggt aggtgtgcgg taggcttttc tccgtcgcag
     3961 gacgcagggt tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg
     4021 aggggaggga taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa
     4081 gtagctgaag ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt
     4141 tttttaggca ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac
     4201 ttgtaaattg tccgctaaat tctggccgtt tttggctttt ttgttagaca acatgggtaa
     4261 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     4321 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     4381 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     4441 tgtttatcgg cactttgcat ccgccgcgct cccgattccg gaagtgcttg acattgggga
     4501 gttcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     4561 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcaa tggatgccat
     4621 cgctgccgcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     4681 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     4741 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     4801 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     4861 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     4921 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     4981 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccccggct
     5041 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     5101 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     5161 gactgtcggg cgtacacaaa tcgcccgcag aagcgccgcc gtctggaccg atggctgtgt
     5221 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     5281 agctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt
     5341 gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag
     5401 tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg
     5461 acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc
     5521 tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg
     5581 caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc
     5641 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5701 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac
     5761 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga
     5821 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5881 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca
     5941 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6001 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa
     6061 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6121 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6181 ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag
     6241 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6301 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6361 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6421 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6481 aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     6541 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     6601 cgtgtcctcc ttaccagaaa tttatcctta agatcgttta aactcgactc tggctctatc
     6661 gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat
     6721 cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct cccgacatct
     6781 tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc agcttcagct
     6841 acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctatc
     6901 aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg atagcgattt
     6961 tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg ggtcattagt
     7021 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     7081 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     7141 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     7201 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     7261 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     7321 ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca tcaatgggcg
     7381 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     7441 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     7501 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
     7561 gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc gccagcggcc
     7621 gc
//
