LOCUS       sequence-295962-        5289 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 5289)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5289)
  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-m4134358
FEATURES             Location/Qualifiers
     source          1..5289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1040..1060
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1061..1105
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1158..1279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1169..1188)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1223..1242
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1394..1440
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1470..1606
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1651..1722
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1948..2536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2028..2047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3282..3350,3351..4142)
                     /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(3500..3519)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4155..4291
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4443..4514
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4519..4565)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4583..4631
                     /note="synthetic polyadenylation signal"
     enhancer        4712..5015
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5017..5219
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5170..5190
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtta gaaaatgtgg cttttaatgt tgtaaataag ggacactttg atggacaaca
       61 gggtgaagta ccagtttcta tcattaataa cactgtttac acaaaagttg atggtgttga
      121 tgtagaattg tttgaaaata aaacaacatt acctgttaat gtagcatttg agctttgggc
      181 taagcgcaac attaaaccag taccagaggt gaaaatactc aataatttgg gtgtggacat
      241 tgctgctaat actgtgatct gggactacaa aagagatgct ccagcacata tatctactat
      301 tggtgtttgt tctatgactg acatagccaa gaaaccaact gaaacgattt gtgcaccact
      361 cactgtcttt tttgatggta gagttgatgg tcaagtagac ttatttagaa atgcccgtaa
      421 tggtgttctt attacagaag gtagtgttaa aggtttacaa ccatctgtag gtcccaaaca
      481 agctagtctt aatggagtca cattaattgg agaagccgta aaaacacagt tcaattatta
      541 taagaaagtt gatggtgttg tccaacaatt acctgaaact tactttactc agagtagaaa
      601 tttacaagaa tttaaaccca ggagtcaaat ggaaattgat ttcttagaat tagctatgga
      661 tgaattcatt gaacggtata aattagaagg ctatgccttc gaacatatcg tttatggaga
      721 ttttagtcat agtcagttag gtggtttaca tctactgatt ggactagcta aacgttttaa
      781 ggaatcacct tttgaattag aagattttat tcctatggac agtacagtta aaaactattt
      841 cataacagat gcgcaaacag gttcatctaa gtgtgtgtgt tctgttattg atttattact
      901 tgatgatttt gttgaaataa taaaatccca agatttatct gtagtttcta aggttgtcaa
      961 agtgactatt gactatacag aaatttcatt tatgctttgg tgtaaagatg gccatgtaga
     1021 aacattttac ccaaaattag aaaaccttta ctttcagtcg aaagaaaccg cggcagctaa
     1081 gttcgagagg cagcacatgg acagttaagc tagcttgact gactgagata cagcgtacct
     1141 tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     1201 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     1261 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     1321 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc
     1381 tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     1441 tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga
     1501 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     1561 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc
     1621 ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca
     1681 ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg
     1741 ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg
     1801 tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg
     1861 ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa
     1921 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     1981 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2041 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2101 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2161 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2221 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2281 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2341 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2401 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2461 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2521 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2581 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2641 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2701 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     2761 atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac
     2821 ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc
     2881 ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga
     2941 atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc
     3001 cgagatccga ggaatatcga aatcggggcg cgcctggtgt accgagaacg atcctctcag
     3061 tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg
     3121 acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc
     3181 tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg
     3241 caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc
     3301 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3361 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac
     3421 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga
     3481 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     3541 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca
     3601 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3661 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa
     3721 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3781 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3841 ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag
     3901 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3961 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4021 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4081 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4141 aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     4201 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     4261 cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg
     4321 ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt
     4381 cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc
     4441 gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc
     4501 ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta
     4561 tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata
     4621 gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt
     4681 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     4741 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     4801 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     4861 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     4921 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     4981 agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca
     5041 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     5101 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     5161 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     5221 gtttagtgaa ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc
     5281 agcggccgc
//
