LOCUS       sequence-295348-        5640 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 5640)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5640)
  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-m4133169
FEATURES             Location/Qualifiers
     source          1..5640
                     /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             1415..1432
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1509..1630
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1520..1539)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1574..1593
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1745..1791
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1821..1957
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2002..2073
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2299..2887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2379..2398)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3633..3701,3702..4493)
                     /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(3851..3870)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4506..4642
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4794..4865
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4870..4916)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4934..4982
                     /note="synthetic polyadenylation signal"
     enhancer        5063..5366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5368..5570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5521..5541
                     /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 cagtcgatga gtgacaaaat
     1081 cattcatcta accgatgact cctttgatac agacgtgttg aaggccgatg gtgcgatact
     1141 ggtagatttc tgggctgaat ggtgtggccc gtgcaagatg atcgcaccca ttctggacga
     1201 gatcgccgac gagtatcaag gaaaattaac tgtcgctaag cttaatatag atcagaaccc
     1261 agggacggca cctaaatacg gcatcagggg aatacctacc ctcctgctct ttaagaatgg
     1321 ggaagttgcc gcgacaaaag tgggagccct gtctaagggc cagctaaaag agttcttgga
     1381 cgctaacctg gcaggttcag gctcggggca catgcaccat caccatcacc atagcagcgg
     1441 gctcgtgcca cgcggctaag ctagcttgac tgactgagat acagcgtacc ttcagctcac
     1501 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     1561 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     1621 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     1681 ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt
     1741 atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg
     1801 ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     1861 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     1921 tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg
     1981 atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc
     2041 agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga
     2101 aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta
     2161 cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat
     2221 tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2281 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2341 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2401 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2461 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2521 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2581 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2641 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2701 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2761 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2821 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2881 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2941 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3001 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3061 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3121 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc
     3181 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg
     3241 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag
     3301 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg
     3361 aggaatatcg aaatcggggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     3421 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     3481 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     3541 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     3601 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     3661 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3721 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     3781 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     3841 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3901 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     3961 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4021 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     4081 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4141 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     4201 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     4261 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4321 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4381 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4441 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     4501 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     4561 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     4621 cttaccagaa atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga
     4681 atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg
     4741 aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg
     4801 gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac
     4861 ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa
     4921 tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta
     4981 ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc
     5041 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     5101 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     5161 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     5221 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     5281 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     5341 acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg
     5401 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     5461 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     5521 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     5581 accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
