LOCUS       sequence-319545-        5277 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 5277)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5277)
  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-m4134331
FEATURES             Location/Qualifiers
     source          1..5277
                     /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             1624..1644
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1645..1671
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1724..1845
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1735..1754)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1789..1808
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1960..2006
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2036..2172
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2217..2288
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2514..3102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2594..2613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3848..3916,3917..4708)
                     /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(4066..4085)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4721..4857
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5009..5080
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5085..5131)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5149..5197
                     /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 ccatgtcact ggagaacgtt
      601 gctttcaatg tagtgaacaa ggggcacttt gacggacagc aaggcgaagt gccggtcagt
      661 atcataaata atacggtgta caccaaagtc gatggtgttg acgtggagtt atttgaaaac
      721 aagacaactt tgcctgtcaa tgtggccttc gagctgtggg caaaacgcaa cattaagcca
      781 gtacccgaag tgaaaatcct caacaatctt ggcgttgata ttgccgcaaa cactgttatc
      841 tgggattata agcgagatgc gcctgcccat atatccacca ttggagtgtg ttctatgaca
      901 gacatcgcta aaaagcccac cgagacaatt tgcgcgccac tcacggtatt ctttgacggg
      961 agggtcgacg gtcaggtgga tctgtttaga aatgcccgta acggcgtact tattactgaa
     1021 ggaagcgtga aaggcctaca gcccagtgtt gggccgaagc aagcaagcct aaatggagtc
     1081 acactgatcg gagaggctgt gaaaacccag ttcaactact ataagaaggt cgacggcgtg
     1141 gttcagcagc tgcctgagac atactttacc caatcgcgga atctgcaaga atttaagccc
     1201 agatcccaga tggagattga cttcttggaa ctggccatgg atgagttcat cgaacggtat
     1261 aagctcgagg gctacgcctt cgagcacata gtttatgggg atttttctca ttcacagtta
     1321 ggcggtctgc acttgctcat cggattagcg aaacgcttta aagagagccc attcgaactt
     1381 gaagacttta tcccaatgga ctcaactgtg aagaattact tcataactga cgctcagaca
     1441 gggtccagca agtgcgtctg ttctgttatt gatttgctac ttgatgattt cgtggagatt
     1501 attaaatcac aggacctgtc cgtggtttcg aaggtcgtga aagtaaccat cgactatacg
     1561 gaaattagtt tcatgctttg gtgtaaggat ggccatgtgg agacatttta tcctaagctc
     1621 caggaaaacc tttactttca gtcgtacccc tatgacgtac cagattatgc gtaagctagc
     1681 ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata cattgatgag
     1741 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     1801 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     1861 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     1921 ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc cttggggcct
     1981 ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc taccggcatt
     2041 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     2101 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     2161 atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga tcaagacatt
     2221 cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca aactttcttc
     2281 cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag tcaagtcagc
     2341 tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt cagtcaagtt
     2401 cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta aatcatgtga
     2461 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2521 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2581 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2641 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2701 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2761 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2821 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2881 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2941 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3001 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3061 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3121 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3181 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3241 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3301 atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc gaactctcca
     3361 aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg
     3421 aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt ggcagcgcct
     3481 atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga gagaagttca
     3541 acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc
     3601 tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg ttgcttggca
     3661 gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat attgtactca
     3721 agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt ctgatcggtc
     3781 aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat cagcaggagg
     3841 ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3901 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3961 gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4021 tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4081 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4141 tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4201 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4261 aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4321 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4381 cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg aactacttac
     4441 tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg caggaccact
     4501 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4561 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4621 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4681 aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc agaaaaaaat
     4741 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     4801 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaagat
     4861 cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg
     4921 aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta
     4981 cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc
     5041 ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag
     5101 acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac
     5161 caacacacat ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag
     5221 ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccg
//
