LOCUS       sequence-295422-        7189 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 7189)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7189)
  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-m4133162
FEATURES             Location/Qualifiers
     source          1..7189
                     /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             1061..1105
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1158..1279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1169..1188)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1223..1242
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1394..1440
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1470..1606
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1651..1722
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1948..2536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2028..2047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3036..4245
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4082..4101
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4249..4848
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4249..4268)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4745..4765
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4875..4923
                     /note="synthetic polyadenylation signal"
     CDS             join(5189..5257,5258..6049)
                     /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(5407..5426)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6062..6198
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6343..6414
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6419..6465)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6483..6531
                     /note="synthetic polyadenylation signal"
     enhancer        6612..6915
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6917..7119
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7070..7090
                     /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 aaagaaaccg cggcagctaa
     1081 gttcgagagg cagcacatgg acagttaagc tagcttgact gactgagata cagcgtacct
     1141 tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     1201 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     1261 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     1321 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc
     1381 tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     1441 tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga
     1501 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     1561 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc
     1621 ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca
     1681 ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg
     1741 ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg
     1801 tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg
     1861 ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa
     1921 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     1981 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2041 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2101 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2161 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2221 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2281 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2341 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2401 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2461 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2521 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2581 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2641 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2701 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     2761 atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac
     2821 ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc
     2881 ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga
     2941 atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc
     3001 cgagatccga ggaatatcga aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc
     3061 tcccgcgggc gcccccctcg tcacggcgag cgctgccacg tcagacgaag ggcgcaggag
     3121 cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc
     3181 ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca
     3241 ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct
     3301 gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg
     3361 gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt
     3421 gatcgtcact tggtgagtag cgggctgctg ggctggccgg ggctttcgtg gccgccgggc
     3481 cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag
     3541 caaggttgcc ctgaactggg ggttgggggg agcgcagcaa aatggcggct gttcccgagt
     3601 cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat
     3661 ggtgggcggc aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg
     3721 ggtgagatgg gctgggcacc atctggggac cctgacgtga agtttgtcac tgactggaga
     3781 actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg gcagtgcacc
     3841 cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat
     3901 aatgcagggt ggggccacct gccggtaggt gtgcggtagg cttttctccg tcgcaggacg
     3961 cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct ctggtgaggg
     4021 gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct tcttaagtag
     4081 ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg tgaagttttt
     4141 taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg ttagacttgt
     4201 aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacaacat gaccgagtac
     4261 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     4321 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     4381 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     4441 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     4501 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     4561 gcgcagcaac agatggaggg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     4621 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     4681 ctccccggag tggaggccgc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     4741 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     4801 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctaggc tagtatgtaa
     4861 gcctagtctt agataataaa atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt
     4921 gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac
     4981 gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc caggaagtcc
     5041 aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt ttaaacctag
     5101 atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa acatctatca
     5161 agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg tcgcccttat
     5221 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5281 aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg atctcaacag
     5341 cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga gcacttttaa
     5401 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5461 ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag aaaagcatct
     5521 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5581 tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg cttttttgca
     5641 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5701 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact
     5761 attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact ggatggaggc
     5821 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5881 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5941 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6001 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac cgattctagg
     6061 tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg
     6121 ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta
     6181 ccagaaattt atccttaaga tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt
     6241 ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag
     6301 ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct
     6361 ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca
     6421 aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta
     6481 cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg
     6541 ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca tagcccatat
     6601 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     6661 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     6721 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     6781 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     6841 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     6901 atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     6961 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     7021 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     7081 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
     7141 agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgc
//
