LOCUS       sequence-297947-        5271 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 5271)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5271)
  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-m4133202
FEATURES             Location/Qualifiers
     source          1..5271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    32..103
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(108..154)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    172..220
                     /note="synthetic polyadenylation signal"
     enhancer        301..604
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        606..808
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     759..779
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1918..1938
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1939..1965
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2018..2139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2029..2048)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2083..2102
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2254..2300
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2330..2466
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2511..2582
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2808..3396)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2888..2907)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4142..4210,4211..5002)
                     /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(4360..4379)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5015..5151
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc
       61 ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc
      121 aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa
      181 aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg
      241 tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      301 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      361 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      421 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      481 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      541 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      601 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      661 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      721 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      781 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      841 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgttaga aaatgtggct
      901 tttaatgttg taaataaggg acactttgat ggacaacagg gtgaagtacc agtttctatc
      961 attaataaca ctgtttacac aaaagttgat ggtgttgatg tagaattgtt tgaaaataaa
     1021 acaacattac ctgttaatgt agcatttgag ctttgggcta agcgcaacat taaaccagta
     1081 ccagaggtga aaatactcaa taatttgggt gtggacattg ctgctaatac tgtgatctgg
     1141 gactacaaaa gagatgctcc agcacatata tctactattg gtgtttgttc tatgactgac
     1201 atagccaaga aaccaactga aacgatttgt gcaccactca ctgtcttttt tgatggtaga
     1261 gttgatggtc aagtagactt atttagaaat gcccgtaatg gtgttcttat tacagaaggt
     1321 agtgttaaag gtttacaacc atctgtaggt cccaaacaag ctagtcttaa tggagtcaca
     1381 ttaattggag aagccgtaaa aacacagttc aattattata agaaagttga tggtgttgtc
     1441 caacaattac ctgaaactta ctttactcag agtagaaatt tacaagaatt taaacccagg
     1501 agtcaaatgg aaattgattt cttagaatta gctatggatg aattcattga acggtataaa
     1561 ttagaaggct atgccttcga acatatcgtt tatggagatt ttagtcatag tcagttaggt
     1621 ggtttacatc tactgattgg actagctaaa cgttttaagg aatcaccttt tgaattagaa
     1681 gattttattc ctatggacag tacagttaaa aactatttca taacagatgc gcaaacaggt
     1741 tcatctaagt gtgtgtgttc tgttattgat ttattacttg atgattttgt tgaaataata
     1801 aaatcccaag atttatctgt agtttctaag gttgtcaaag tgactattga ctatacagaa
     1861 atttcattta tgctttggtg taaagatggc catgtagaaa cattttaccc aaaattagaa
     1921 aacctttact ttcagtcgta cccctatgac gtaccagatt atgcgtaagc tagcttgact
     1981 gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga
     2041 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2101 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2161 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     2221 aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac
     2281 gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca
     2341 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     2401 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     2461 ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta
     2521 atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc
     2581 taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg
     2641 gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct
     2701 ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa
     2761 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2821 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2881 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2941 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3001 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3061 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3121 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3181 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3241 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3301 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3361 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3421 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3481 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3541 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3601 gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc
     3661 tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa
     3721 ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc
     3781 aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac
     3841 atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggtgt
     3901 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
     3961 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
     4021 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
     4081 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
     4141 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4201 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4261 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4321 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4381 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4441 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4501 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4561 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4621 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4681 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
     4741 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4801 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4861 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4921 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4981 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
     5041 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     5101 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa
     5161 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     5221 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag c
//
