LOCUS       sequence-295522-        7171 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 7171)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7171)
  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-m4133944
FEATURES             Location/Qualifiers
     source          1..7171
                     /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..1087
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1140..1261
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1151..1170)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1205..1224
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1376..1422
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1452..1588
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1633..1704
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1930..2518)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2010..2029)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3018..4227
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4064..4083
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4231..4830
                     /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(4231..4250)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4727..4747
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4857..4905
                     /note="synthetic polyadenylation signal"
     CDS             join(5171..5239,5240..6031)
                     /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(5389..5408)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6044..6180
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6325..6396
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6401..6447)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6465..6513
                     /note="synthetic polyadenylation signal"
     enhancer        6594..6897
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6899..7101
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7052..7072
                     /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 tacccctatg acgtaccaga
     1081 ttatgcgtaa gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat
     1141 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1201 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1261 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1321 ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca
     1381 taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt
     1441 gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     1501 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     1561 gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga
     1621 tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt
     1681 tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt
     1741 aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa
     1801 ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt
     1861 catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1921 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1981 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2041 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2101 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2161 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2221 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2281 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2341 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2401 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2461 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2521 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2581 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2641 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2701 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa
     2761 atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct
     2821 tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta
     2881 ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc
     2941 acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc
     3001 gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
     3061 cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga tccttccgcc
     3121 cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag
     3181 cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag
     3241 agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg
     3301 ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg
     3361 cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt
     3421 agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag
     3481 cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg
     3541 ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt
     3601 gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc
     3661 aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat gggctgggca
     3721 ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt
     3781 tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg
     3841 cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac
     3901 ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg
     3961 taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt
     4021 cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga
     4081 actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg
     4141 taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc cgctaaattc
     4201 tggccgtttt tggctttttt gttagacaac atgaccgagt acaagcccac ggtgcgcctc
     4261 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     4321 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     4381 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     4441 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     4501 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggag
     4561 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     4621 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcc
     4681 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     4741 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     4801 tggtgcatga cccgcaagcc cggtgcctag gctagtatgt aagcctagtc ttagataata
     4861 aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc
     4921 gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata tcgttgcttg
     4981 gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca gatattgtac
     5041 tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat agtctgatcg
     5101 gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag gatcagcagg
     5161 aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5221 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5281 gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5341 ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5401 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5461 gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg gcatgacagt
     5521 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5581 gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5641 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5701 caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg gcgaactact
     5761 tactctagct tcccggcaac agttgataga ctggatggag gcggataaag ttgcaggacc
     5821 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5881 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5941 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6001 gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg cgcagaaaaa
     6061 aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat
     6121 acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa
     6181 gatcgtttaa actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca
     6241 gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt
     6301 tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc
     6361 ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc
     6421 gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac
     6481 acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag
     6541 atctagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     6601 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     6661 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     6721 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     6781 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     6841 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg
     6901 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     6961 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     7021 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     7081 gaggtctata taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta
     7141 ccatccactc gacacacccg ccagcggccg c
//
