LOCUS       sequence-295261-        5328 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 5328)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5328)
  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-m4134170
FEATURES             Location/Qualifiers
     source          1..5328
                     /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             1067..1144
                     /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"
     polyA_signal    1197..1318
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1208..1227)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1262..1281
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1433..1479
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1509..1645
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1690..1761
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1987..2575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2067..2086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3321..3389,3390..4181)
                     /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(3539..3558)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4194..4330
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4482..4553
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4558..4604)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4622..4670
                     /note="synthetic polyadenylation signal"
     enhancer        4751..5054
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5056..5258
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5209..5229
                     /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 cagtcgaaga ggcgatggaa
     1081 aaagaatttt atcgcggtga gtgcagccaa cagattcaaa aagatatcct cgtctggtgc
     1141 cctctaagct agcttgactg actgagatac agcgtacctt cagctcacag acatgataag
     1201 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     1261 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     1321 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     1381 aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa
     1441 ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct
     1501 atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     1561 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     1621 ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt
     1681 cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca
     1741 tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac
     1801 cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg
     1861 cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat
     1921 ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1981 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2041 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2101 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2161 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2221 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2281 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2341 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2401 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2461 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2521 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2581 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2641 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2701 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2761 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt
     2821 tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc
     2881 aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg
     2941 cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc
     3001 cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa
     3061 atcggggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     3121 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     3181 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     3241 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     3301 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     3361 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3421 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3481 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     3541 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3601 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     3661 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3721 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3781 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3841 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     3901 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     3961 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4021 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4081 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4141 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     4201 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     4261 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     4321 ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt
     4381 tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc
     4441 tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc
     4501 cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa
     4561 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac
     4621 acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc
     4681 tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata
     4741 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     4801 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     4861 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     4921 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     4981 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     5041 tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     5101 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     5161 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     5221 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca
     5281 gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgc
//
