LOCUS       sequence-295102-        7168 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 7168)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7168)
  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-m4133322
FEATURES             Location/Qualifiers
     source          1..7168
                     /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..1084
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1137..1258
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1148..1167)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1202..1221
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1373..1419
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1449..1585
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1630..1701
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1927..2515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2007..2026)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3015..4224
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4061..4080
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4228..4827
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4228..4247)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4724..4744
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4854..4902
                     /note="synthetic polyadenylation signal"
     CDS             join(5168..5236,5237..6028)
                     /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(5386..5405)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6041..6177
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6322..6393
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6398..6444)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6462..6510
                     /note="synthetic polyadenylation signal"
     enhancer        6591..6894
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6896..7098
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7049..7069
                     /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 cagtcgcacc atcaccatca
     1081 ccattaagct agcttgactg actgagatac agcgtacctt cagctcacag acatgataag
     1141 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     1201 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     1261 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     1321 aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa
     1381 ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct
     1441 atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     1501 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     1561 ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt
     1621 cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca
     1681 tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac
     1741 cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg
     1801 cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat
     1861 ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1921 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1981 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2041 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2101 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2161 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2221 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2281 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2341 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2401 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2461 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2521 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2581 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2641 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2701 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt
     2761 tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc
     2821 aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg
     2881 cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc
     2941 cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa
     3001 atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt
     3061 cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg
     3121 acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag
     3181 aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc
     3241 ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc
     3301 ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag
     3361 ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc
     3421 gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt
     3481 gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg
     3541 gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg
     3601 aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag
     3661 gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctgggcacca
     3721 tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc
     3781 gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc
     3841 cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccacctg
     3901 ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag
     3961 gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag
     4021 tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact
     4081 atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa
     4141 tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc taaattctgg
     4201 ccgtttttgg cttttttgtt agacaacatg accgagtaca agcccacggt gcgcctcgcc
     4261 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     4321 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     4381 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     4441 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     4501 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggagggc
     4561 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     4621 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggccgcc
     4681 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     4741 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     4801 tgcatgaccc gcaagcccgg tgcctaggct agtatgtaag cctagtctta gataataaaa
     4861 tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc
     4921 tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg ttgcttggca
     4981 gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat attgtactca
     5041 agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt ctgatcggtc
     5101 aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat cagcaggagg
     5161 ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5221 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5281 gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5341 tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5401 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5461 tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5521 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5581 aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5641 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5701 cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg aactacttac
     5761 tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg caggaccact
     5821 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5881 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5941 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6001 aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc agaaaaaaat
     6061 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     6121 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaagat
     6181 cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc
     6241 gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt
     6301 ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc
     6361 tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt
     6421 tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca
     6481 catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc
     6541 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     6601 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     6661 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     6721 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     6781 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     6841 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg
     6901 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     6961 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     7021 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     7081 gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca
     7141 tccactcgac acacccgcca gcggccgc
//
