LOCUS       sequence-297882-        7665 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 7665)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7665)
  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-m4134257
FEATURES             Location/Qualifiers
     source          1..7665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1314..1334
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1338..1403
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1472..1593
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1483..1502)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1537..1556
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1708..1754
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1784..1920
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1965..2036
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2262..2850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2342..2361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3350..4559
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4396..4415
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4563..5591
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5618..5666
                     /note="synthetic polyadenylation signal"
     CDS             join(5932..6000,6001..6792)
                     /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(6150..6169)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6805..6941
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7093..7164
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7169..7215)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7233..7281
                     /note="synthetic polyadenylation signal"
     enhancer        7362..7665
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc
      241 ctaccatcca ctcgacacac ccgccagcgg ccgccaccat gttagaaaat gtggctttta
      301 atgttgtaaa taagggacac tttgatggac aacagggtga agtaccagtt tctatcatta
      361 ataacactgt ttacacaaaa gttgatggtg ttgatgtaga attgtttgaa aataaaacaa
      421 cattacctgt taatgtagca tttgagcttt gggctaagcg caacattaaa ccagtaccag
      481 aggtgaaaat actcaataat ttgggtgtgg acattgctgc taatactgtg atctgggact
      541 acaaaagaga tgctccagca catatatcta ctattggtgt ttgttctatg actgacatag
      601 ccaagaaacc aactgaaacg atttgtgcac cactcactgt cttttttgat ggtagagttg
      661 atggtcaagt agacttattt agaaatgccc gtaatggtgt tcttattaca gaaggtagtg
      721 ttaaaggttt acaaccatct gtaggtccca aacaagctag tcttaatgga gtcacattaa
      781 ttggagaagc cgtaaaaaca cagttcaatt attataagaa agttgatggt gttgtccaac
      841 aattacctga aacttacttt actcagagta gaaatttaca agaatttaaa cccaggagtc
      901 aaatggaaat tgatttctta gaattagcta tggatgaatt cattgaacgg tataaattag
      961 aaggctatgc cttcgaacat atcgtttatg gagattttag tcatagtcag ttaggtggtt
     1021 tacatctact gattggacta gctaaacgtt ttaaggaatc accttttgaa ttagaagatt
     1081 ttattcctat ggacagtaca gttaaaaact atttcataac agatgcgcaa acaggttcat
     1141 ctaagtgtgt gtgttctgtt attgatttat tacttgatga ttttgttgaa ataataaaat
     1201 cccaagattt atctgtagtt tctaaggttg tcaaagtgac tattgactat acagaaattt
     1261 catttatgct ttggtgtaaa gatggccatg tagaaacatt ttacccaaaa ttagaaaacc
     1321 tttactttca gtcgatggat tataaggacc acgatggaga ctacaaagat catgacattg
     1381 attacaagga cgacgatgac aaataaggat ccgcaggcct ctgctagctt gactgactga
     1441 gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt tggacaaacc
     1501 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     1561 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     1621 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     1681 ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct aaacgggtct
     1741 tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa
     1801 aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat
     1861 acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa
     1921 ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc
     1981 ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct
     2041 cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga
     2101 tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg atctggtcct
     2161 tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc aaaaggccag
     2221 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2281 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2341 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2401 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2461 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2521 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2581 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2641 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2701 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2761 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2821 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2881 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2941 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3001 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3061 tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag gccctcgtcg
     3121 gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa cgaactatcg
     3181 caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat cgccaggtat
     3241 tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac ctacatcctc
     3301 aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc
     3361 cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac
     3421 gaagggcgca ggagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct
     3481 cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg
     3541 gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc
     3601 ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg
     3661 acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt
     3721 ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt
     3781 cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga gagaccgcca agggctgtag
     3841 tctgggtccg cgagcaaggt tgccctgaac tgggggttgg ggggagcgca gcaaaatggc
     3901 ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac
     3961 aaggtggggg gcatggtggg cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg
     4021 aaagctctta ttcgggtgag atgggctggg caccatctgg ggaccctgac gtgaagtttg
     4081 tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat ggcggtgccg
     4141 ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg
     4201 ttctgttggc ttataatgca gggtggggcc acctgccggt aggtgtgcgg taggcttttc
     4261 tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga caggcgccgg
     4321 acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt ttatgtacct
     4381 atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt
     4441 tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc
     4501 agtgttagac ttgtaaattg tccgctaaat tctggccgtt tttggctttt ttgttagaca
     4561 acatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     4621 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     4681 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     4741 aagatcgtta tgtttatcgg cactttgcat ccgccgcgct cccgattccg gaagtgcttg
     4801 acattgggga gttcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     4861 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcaa
     4921 tggatgccat cgctgccgcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     4981 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     5041 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     5101 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     5161 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     5221 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     5281 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     5341 cgccccggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     5401 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     5461 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgccgcc gtctggaccg
     5521 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     5581 caaaggaata agctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag
     5641 atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg
     5701 atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat
     5761 ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag
     5821 cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc
     5881 ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt
     5941 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6001 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt
     6061 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc
     6121 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6181 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat
     6241 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6301 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg
     6361 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6421 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6481 atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6541 cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6601 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6661 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6721 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6781 aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg
     6841 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
     6901 cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac
     6961 tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc
     7021 atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat
     7081 cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca
     7141 taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc
     7201 caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct
     7261 tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca
     7321 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     7381 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     7441 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     7501 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     7561 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     7621 cctacttggc agtacatcta cgtattagtc atcgctatta ccatg
//
