LOCUS       sequence-307289-        7169 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 7169)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7169)
  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-m4133166
FEATURES             Location/Qualifiers
     source          1..7169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        81..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     539..559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1698..1718
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1719..1736
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1789..1910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1800..1819)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1854..1873
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2025..2071
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2101..2237
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2282..2353
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2579..3167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2659..2678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3667..4876
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4713..4732
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4880..5479
                     /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(4880..4899)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5376..5396
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5506..5554
                     /note="synthetic polyadenylation signal"
     CDS             join(5820..5888,5889..6680)
                     /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(6038..6057)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6693..6829
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6981..7052
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7057..7103)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7121..7169
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga
      421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
      601 cgtcagatca gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgcca
      661 ccatgttaga aaatgtggct tttaatgttg taaataaggg acactttgat ggacaacagg
      721 gtgaagtacc agtttctatc attaataaca ctgtttacac aaaagttgat ggtgttgatg
      781 tagaattgtt tgaaaataaa acaacattac ctgttaatgt agcatttgag ctttgggcta
      841 agcgcaacat taaaccagta ccagaggtga aaatactcaa taatttgggt gtggacattg
      901 ctgctaatac tgtgatctgg gactacaaaa gagatgctcc agcacatata tctactattg
      961 gtgtttgttc tatgactgac atagccaaga aaccaactga aacgatttgt gcaccactca
     1021 ctgtcttttt tgatggtaga gttgatggtc aagtagactt atttagaaat gcccgtaatg
     1081 gtgttcttat tacagaaggt agtgttaaag gtttacaacc atctgtaggt cccaaacaag
     1141 ctagtcttaa tggagtcaca ttaattggag aagccgtaaa aacacagttc aattattata
     1201 agaaagttga tggtgttgtc caacaattac ctgaaactta ctttactcag agtagaaatt
     1261 tacaagaatt taaacccagg agtcaaatgg aaattgattt cttagaatta gctatggatg
     1321 aattcattga acggtataaa ttagaaggct atgccttcga acatatcgtt tatggagatt
     1381 ttagtcatag tcagttaggt ggtttacatc tactgattgg actagctaaa cgttttaagg
     1441 aatcaccttt tgaattagaa gattttattc ctatggacag tacagttaaa aactatttca
     1501 taacagatgc gcaaacaggt tcatctaagt gtgtgtgttc tgttattgat ttattacttg
     1561 atgattttgt tgaaataata aaatcccaag atttatctgt agtttctaag gttgtcaaag
     1621 tgactattga ctatacagaa atttcattta tgctttggtg taaagatggc catgtagaaa
     1681 cattttaccc aaaattagaa aacctttact ttcagtcgca ccatcaccat caccattaag
     1741 ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg
     1801 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1861 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1921 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1981 aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg
     2041 ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg
     2101 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     2161 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     2221 cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag
     2281 acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt
     2341 tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag
     2401 tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc
     2461 aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca
     2521 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2581 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2641 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2701 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2761 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2821 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2881 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2941 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3001 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3061 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3121 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3181 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3241 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3301 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3361 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact
     3421 ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc
     3481 tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag
     3541 cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa
     3601 gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc
     3661 gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga
     3721 gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag
     3781 gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat
     3841 tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg
     3901 cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg
     3961 ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt
     4021 tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct
     4081 gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag
     4141 accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg
     4201 gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc
     4261 tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag
     4321 cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga
     4381 ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg
     4441 cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg
     4501 tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg
     4561 tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct
     4621 gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg
     4681 gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc
     4741 ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg
     4801 gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt
     4861 ggcttttttg ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     4921 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     4981 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     5041 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     5101 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     5161 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc
     5221 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     5281 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc
     5341 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     5401 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     5461 ccgcaagccc ggtgcctagg ctagtatgta agcctagtct tagataataa aatcgctatc
     5521 catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac
     5581 cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca
     5641 gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt
     5701 cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata
     5761 atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca
     5821 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5881 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc
     5941 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6001 aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6061 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6121 ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6181 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg
     6241 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6301 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6361 ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt
     6421 cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6481 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6541 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6601 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6661 cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg
     6721 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     6781 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc
     6841 gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg
     6901 tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg
     6961 gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct tcttccctct
     7021 agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt
     7081 agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac
     7141 atccatcttc gatggatagc gattttatt
//
