LOCUS       sequence-295544-        7186 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 7186)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7186)
  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-m4133999
FEATURES             Location/Qualifiers
     source          1..7186
                     /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..1102
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1155..1276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1166..1185)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1220..1239
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1391..1437
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1467..1603
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1648..1719
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1945..2533)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2025..2044)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3033..4242
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4079..4098
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4246..4845
                     /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(4246..4265)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4742..4762
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4872..4920
                     /note="synthetic polyadenylation signal"
     CDS             join(5186..5254,5255..6046)
                     /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(5404..5423)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6059..6195
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6340..6411
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6416..6462)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6480..6528
                     /note="synthetic polyadenylation signal"
     enhancer        6609..6912
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6914..7116
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7067..7087
                     /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 gggaaaccta tacccaatcc
     1081 acttctcggc ttagattcta cctaagctag cttgactgac tgagatacag cgtaccttca
     1141 gctcacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     1201 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1261 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     1321 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct
     1381 atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc
     1441 ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc
     1501 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     1561 cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg
     1621 caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta
     1681 ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc
     1741 tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt
     1801 cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc
     1861 tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1921 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1981 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2041 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2101 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2161 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2221 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2281 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2341 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2401 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2461 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2521 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2581 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2641 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2701 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2761 catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt
     2821 tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc
     2881 cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata
     2941 tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga
     3001 gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc
     3061 cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt
     3121 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta
     3181 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg
     3241 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg
     3301 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca
     3361 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat
     3421 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc
     3481 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa
     3541 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt
     3601 gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt
     3661 gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt
     3721 gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact
     3781 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt
     3841 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat
     3901 gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag
     3961 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag
     4021 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg
     4081 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag
     4141 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa
     4201 ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag
     4261 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     4321 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     4381 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     4441 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     4501 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     4561 cagcaacaga tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     4621 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     4681 cccggagtgg aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     4741 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     4801 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc
     4861 tagtcttaga taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg
     4921 tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat
     4981 ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat
     5041 cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata
     5101 ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga
     5161 gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc
     5221 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5281 agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg
     5341 taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt
     5401 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5461 catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac
     5521 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5581 ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa
     5641 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5701 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt
     5761 aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga
     5821 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5881 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5941 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6001 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc
     6061 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     6121 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     6181 gaaatttatc cttaagatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc
     6241 gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta
     6301 tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca
     6361 caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa
     6421 aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac
     6481 acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg
     6541 atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg
     6601 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     6661 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     6721 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     6781 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     6841 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     6901 gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     6961 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     7021 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     7081 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga
     7141 tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgc
//
