LOCUS       sequence-295926-        7705 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 7705)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7705)
  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-m4134339
FEATURES             Location/Qualifiers
     source          1..7705
                     /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             1100..1177
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             1205..1225
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1674..1795
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1685..1704)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1739..1758
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1910..1956
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1986..2122
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2167..2238
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2464..3052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2544..2563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3552..4761
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4598..4617
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4765..5364
                     /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(4765..4784)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5261..5281
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5391..5439
                     /note="synthetic polyadenylation signal"
     CDS             join(5705..5773,5774..6565)
                     /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(5923..5942)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6578..6714
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6859..6930
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6935..6981)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6999..7047
                     /note="synthetic polyadenylation signal"
     enhancer        7128..7431
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7433..7635
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7586..7606
                     /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 cagtcgggac ggcgaatccc
     1081 ggggctgata aatccctgga aaaggagatg gaagaagaac tttattgcag tgtctgcggc
     1141 taaccggttc aaaaagatca gttcaagcgg cgccctagat tacgacattc ctacgaccgc
     1201 ctccgagaat ctctatttcc agggtgaatt tggcttggca caacatgacg aggccgtcga
     1261 taacaagttt aataaagaac agcagaacgc tttctatgag atccttcact taccaaatct
     1321 gaacgaagag caacgcaacg ctttcattca gagcctgaaa gacgatccct cgcagtctgc
     1381 aaatctcctg gccgaagcca agaagctgaa tgacgcacag gctccaaaag ttgataacaa
     1441 gtttaacaag gagcagcaga atgctttcta cgagatattg caccttccta atctgaatga
     1501 ggaacaaaga aatgcattta tccagtccct aaaagacgat ccatcacaga gtgccaacct
     1561 gcttgcggaa gccaaaaagc tcaatgatgc ccaggccccg aaagtagacg cgaaccatca
     1621 ataagctagc ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata
     1681 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     1741 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     1801 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     1861 caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc
     1921 cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc
     1981 taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     2041 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     2101 cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga
     2161 tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca
     2221 aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag
     2281 tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt
     2341 cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta
     2401 aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2461 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2521 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2581 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2641 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2701 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2761 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2821 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2881 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2941 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3001 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3061 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3121 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3181 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3241 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc
     3301 gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg
     3361 ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt
     3421 ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga
     3481 gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc
     3541 ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac
     3601 ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc cgcccggacg
     3661 ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag
     3721 gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga
     3781 acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt
     3841 gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg
     3901 ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg
     3961 ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg
     4021 gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt
     4081 ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg
     4141 cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc
     4201 ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg ggcaccatct
     4261 ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg
     4321 ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag cgcgcgccct
     4381 cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg ccacctgccg
     4441 gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct agggtaggct
     4501 ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag gcgtcagttt
     4561 ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt ttgaactatg
     4621 cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga aatgtaatca
     4681 tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa attctggccg
     4741 tttttggctt ttttgttaga caacatgacc gagtacaagc ccacggtgcg cctcgccacc
     4801 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     4861 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     4921 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     4981 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     5041 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggagggcctc
     5101 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     5161 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggccgccgag
     5221 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     5281 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     5341 atgacccgca agcccggtgc ctaggctagt atgtaagcct agtcttagat aataaaatcg
     5401 ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg
     5461 tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg cttggcagtc
     5521 agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt gtactcaagc
     5581 ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg atcggtcaac
     5641 gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag caggaggctt
     5701 tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5761 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5821 tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5881 ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5941 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6001 cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6061 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6121 gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6181 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6241 gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct
     6301 agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct
     6361 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6421 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6481 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6541 tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc
     6601 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     6661 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatcgt
     6721 ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg
     6781 gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc
     6841 agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag
     6901 tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag
     6961 aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat
     7021 ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag
     7081 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     7141 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     7201 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     7261 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     7321 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     7381 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg
     7441 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     7501 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     7561 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     7621 tatataagca gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc
     7681 actcgacaca cccgccagcg gccgc
//
