LOCUS       sequence-308743-        7706 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 7706)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7706)
  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-m4133248
FEATURES             Location/Qualifiers
     source          1..7706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1618..1638
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1672..1749
                     /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             1777..1797
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2246..2367
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2257..2276)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2311..2330
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2482..2528
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2558..2694
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2739..2810
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3036..3624)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3116..3135)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4124..5333
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5170..5189
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5337..5936
                     /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(5337..5356)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5833..5853
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5963..6011
                     /note="synthetic polyadenylation signal"
     CDS             join(6277..6345,6346..7137)
                     /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(6495..6514)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7150..7286
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7438..7509
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7514..7560)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7578..7626
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgttaga aaatgtggct
      601 tttaatgttg taaataaggg acactttgat ggacaacagg gtgaagtacc agtttctatc
      661 attaataaca ctgtttacac aaaagttgat ggtgttgatg tagaattgtt tgaaaataaa
      721 acaacattac ctgttaatgt agcatttgag ctttgggcta agcgcaacat taaaccagta
      781 ccagaggtga aaatactcaa taatttgggt gtggacattg ctgctaatac tgtgatctgg
      841 gactacaaaa gagatgctcc agcacatata tctactattg gtgtttgttc tatgactgac
      901 atagccaaga aaccaactga aacgatttgt gcaccactca ctgtcttttt tgatggtaga
      961 gttgatggtc aagtagactt atttagaaat gcccgtaatg gtgttcttat tacagaaggt
     1021 agtgttaaag gtttacaacc atctgtaggt cccaaacaag ctagtcttaa tggagtcaca
     1081 ttaattggag aagccgtaaa aacacagttc aattattata agaaagttga tggtgttgtc
     1141 caacaattac ctgaaactta ctttactcag agtagaaatt tacaagaatt taaacccagg
     1201 agtcaaatgg aaattgattt cttagaatta gctatggatg aattcattga acggtataaa
     1261 ttagaaggct atgccttcga acatatcgtt tatggagatt ttagtcatag tcagttaggt
     1321 ggtttacatc tactgattgg actagctaaa cgttttaagg aatcaccttt tgaattagaa
     1381 gattttattc ctatggacag tacagttaaa aactatttca taacagatgc gcaaacaggt
     1441 tcatctaagt gtgtgtgttc tgttattgat ttattacttg atgattttgt tgaaataata
     1501 aaatcccaag atttatctgt agtttctaag gttgtcaaag tgactattga ctatacagaa
     1561 atttcattta tgctttggtg taaagatggc catgtagaaa cattttaccc aaaattagaa
     1621 aacctttact ttcagtcggg acggcgaatc ccggggctga taaatccctg gaaaaggaga
     1681 tggaagaaga actttattgc agtgtctgcg gctaaccggt tcaaaaagat cagttcaagc
     1741 ggcgccctag attacgacat tcctacgacc gcctccgaga atctctattt ccagggtgaa
     1801 tttggcttgg cacaacatga cgaggccgtc gataacaagt ttaataaaga acagcagaac
     1861 gctttctatg agatccttca cttaccaaat ctgaacgaag agcaacgcaa cgctttcatt
     1921 cagagcctga aagacgatcc ctcgcagtct gcaaatctcc tggccgaagc caagaagctg
     1981 aatgacgcac aggctccaaa agttgataac aagtttaaca aggagcagca gaatgctttc
     2041 tacgagatat tgcaccttcc taatctgaat gaggaacaaa gaaatgcatt tatccagtcc
     2101 ctaaaagacg atccatcaca gagtgccaac ctgcttgcgg aagccaaaaa gctcaatgat
     2161 gcccaggccc cgaaagtaga cgcgaaccat caataagcta gcttgactga ctgagataca
     2221 gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca aaccacaact
     2281 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     2341 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     2401 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt
     2461 ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg
     2521 gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc
     2581 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     2641 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt
     2701 cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct
     2761 ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg
     2821 ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc gagatcgact
     2881 tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat
     2941 tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag
     3001 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3061 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3121 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3181 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3241 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3301 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3361 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3421 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3481 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3541 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3601 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3661 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3721 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3781 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3841 tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc gtcggaaaat
     3901 cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact atcgcaagtc
     3961 tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag gtattactcc
     4021 aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat cctcaatccc
     4081 gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggcctcc gcgccgggtt
     4141 ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc agacgaaggg
     4201 cgcaggagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag
     4261 actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact
     4321 ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg
     4381 gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc
     4441 cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg
     4501 atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc
     4561 cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg
     4621 tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa tggcggctgt
     4681 tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg
     4741 gggggcatgg tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg cgggaaagct
     4801 cttattcggg tgagatgggc tgggcaccat ctggggaccc tgacgtgaag tttgtcactg
     4861 actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc
     4921 agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt
     4981 tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc
     5041 gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct
     5101 ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc
     5161 ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg
     5221 aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt
     5281 agacttgtaa attgtccgct aaattctggc cgtttttggc ttttttgtta gacaacatga
     5341 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     5401 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     5461 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     5521 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     5581 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     5641 ggctggccgc gcagcaacag atggagggcc tcctggcgcc gcaccggccc aaggagcccg
     5701 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     5761 ccgtcgtgct ccccggagtg gaggccgccg agcgcgccgg ggtgcccgcc ttcctggaga
     5821 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     5881 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctaggcta
     5941 gtatgtaagc ctagtcttag ataataaaat cgctatccat cgaagatgga tgtgtgttgg
     6001 ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga
     6061 gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca
     6121 ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt
     6181 aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac
     6241 atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc
     6301 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6361 gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat
     6421 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc
     6481 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6541 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa
     6601 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6661 gataacactg cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct
     6721 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6781 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg
     6841 cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg
     6901 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6961 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     7021 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     7081 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg
     7141 attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc
     7201 cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt
     7261 cctccttacc agaaatttat ccttaagatc ccgaatcgtt taaactcgac tctggctcta
     7321 tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc
     7381 atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat
     7441 cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag
     7501 ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta
     7561 tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat
     7621 tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta
     7681 gttcatagcc catatatgga gttccg
//
