LOCUS       sequence-295260-        7979 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 7979)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7979)
  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-m4134141
FEATURES             Location/Qualifiers
     source          1..7979
                     /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             1409..1426
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1519..1640
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1530..1549)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1584..1603
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1755..1801
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1831..1967
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2012..2083
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2309..2897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2389..2408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3397..4606
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4443..4462
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4610..5638
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5665..5713
                     /note="synthetic polyadenylation signal"
     CDS             join(5979..6047,6048..6839)
                     /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(6197..6216)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6852..6988
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7133..7204
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7209..7255)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7273..7321
                     /note="synthetic polyadenylation signal"
     enhancer        7402..7705
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7707..7909
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7860..7880
                     /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 atgagtgaca aaatcattca
     1081 tctaaccgat gactcctttg atacagacgt gttgaaggcc gatggtgcga tactggtaga
     1141 tttctgggct gaatggtgtg gcccgtgcaa gatgatcgca cccattctgg acgagatcgc
     1201 cgacgagtat caaggaaaat taactgtcgc taagcttaat atagatcaga acccagggac
     1261 ggcacctaaa tacggcatca ggggaatacc taccctcctg ctctttaaga atggggaagt
     1321 tgccgcgaca aaagtgggag ccctgtctaa gggccagcta aaagagttct tggacgctaa
     1381 cctggcaggt tcaggctcgg ggcacatgca ccatcaccat caccatagca gcgggctcgt
     1441 gccacgcggc taaggatccg caggcctctg ctagcttgac tgactgagat acagcgtacc
     1501 ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     1561 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     1621 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     1681 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat
     1741 ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1801 gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg
     1861 acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa
     1921 cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat
     1981 cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc
     2041 actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt
     2101 gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg
     2161 gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc
     2221 gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga
     2281 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2341 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2401 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2461 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2521 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2581 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2641 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2701 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2761 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2821 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2881 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2941 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3001 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3061 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3121 catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa
     3181 cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc
     3241 cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg
     3301 aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat
     3361 ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc
     3421 ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga
     3481 gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc
     3541 cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc
     3601 actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc
     3661 tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt
     3721 ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg
     3781 tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg
     3841 ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga
     3901 gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag
     3961 tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca
     4021 tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc
     4081 gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag
     4141 aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac
     4201 ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta
     4261 taatgcaggg tggggccacc tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac
     4321 gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg
     4381 ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta
     4441 gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt
     4501 ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg
     4561 taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca tgggtaaaaa
     4621 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     4681 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     4741 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     4801 ttatcggcac tttgcatccg ccgcgctccc gattccggaa gtgcttgaca ttggggagtt
     4861 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     4921 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggcaatgg atgccatcgc
     4981 tgccgccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     5041 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     5101 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     5161 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     5221 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     5281 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     5341 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cccggctccg
     5401 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     5461 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     5521 tgtcgggcgt acacaaatcg cccgcagaag cgccgccgtc tggaccgatg gctgtgtaga
     5581 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaataagc
     5641 tagtatgtaa gcctagtctt agataataaa atcgctatcc atcgaagatg gatgtgtgtt
     5701 ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc
     5761 gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc
     5821 caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt
     5881 ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa
     5941 acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg
     6001 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6061 tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg
     6121 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga
     6181 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6241 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag
     6301 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6361 gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg
     6421 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6481 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct
     6541 tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact
     6601 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6661 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6721 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6781 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6841 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     6901 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     6961 gtcctcctta ccagaaattt atccttaaga tcgtttaaac tcgactctgg ctctatcgaa
     7021 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
     7081 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
     7141 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
     7201 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat
     7261 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat
     7321 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca
     7381 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     7441 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     7501 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     7561 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     7621 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     7681 cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg
     7741 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     7801 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     7861 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     7921 ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgc
//
