LOCUS       sequence-295250-        7195 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 7195)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7195)
  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-m4134057
FEATURES             Location/Qualifiers
     source          1..7195
                     /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..1111
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1164..1285
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1175..1194)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1229..1248
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1400..1446
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1476..1612
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1657..1728
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1954..2542)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2034..2053)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3042..4251
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4088..4107
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4255..4854
                     /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(4255..4274)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4751..4771
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4881..4929
                     /note="synthetic polyadenylation signal"
     CDS             join(5195..5263,5264..6055)
                     /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(5413..5432)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6068..6204
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6349..6420
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6425..6471)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6489..6537
                     /note="synthetic polyadenylation signal"
     enhancer        6618..6921
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6923..7125
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7076..7096
                     /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 cagtcgaaag aaaccgcggc
     1081 agctaagttc gagaggcagc acatggacag ttaagctagc ttgactgact gagatacagc
     1141 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1201 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1261 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1321 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     1381 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1441 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     1501 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     1561 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     1621 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     1681 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     1741 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     1801 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     1861 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     1921 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1981 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2041 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2101 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2161 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2221 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2281 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2341 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2401 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2461 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2521 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2581 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2641 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2701 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2761 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     2821 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     2881 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     2941 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     3001 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt
     3061 ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg
     3121 caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac
     3181 tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct
     3241 agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc
     3301 gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg
     3361 ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat
     3421 cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg
     3481 ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc
     3541 cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc
     3601 ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg
     3661 gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg ggaaagctct
     3721 tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac
     3781 tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag
     3841 tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg
     3901 gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc
     3961 aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg
     4021 tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt
     4081 aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa
     4141 gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag
     4201 acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga caacatgacc
     4261 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     4321 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     4381 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     4441 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     4501 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     4561 ctggccgcgc agcaacagat ggagggcctc ctggcgccgc accggcccaa ggagcccgcg
     4621 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     4681 gtcgtgctcc ccggagtgga ggccgccgag cgcgccgggg tgcccgcctt cctggagacc
     4741 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     4801 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctaggctagt
     4861 atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg tgtgttggtt
     4921 ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt
     4981 ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt gggacccagg
     5041 aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa
     5101 acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt ttgcaaacat
     5161 ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt tccgtgtcgc
     5221 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5281 gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg aactggatct
     5341 caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa tgatgagcac
     5401 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5461 cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag tcacagaaaa
     5521 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5581 taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc taaccgcttt
     5641 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5701 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caaccttgcg
     5761 taaactatta actggcgaac tacttactct agcttcccgg caacagttga tagactggat
     5821 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5881 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5941 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6001 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaaccgat
     6061 tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     6121 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     6181 tccttaccag aaatttatcc ttaagatcgt ttaaactcga ctctggctct atcgaatctc
     6241 cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct
     6301 cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca
     6361 ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc
     6421 aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt
     6481 gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc
     6541 taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc
     6601 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     6661 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     6721 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     6781 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     6841 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     6901 ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag
     6961 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     7021 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     7081 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
     7141 cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgc
//
