LOCUS       sequence-317257-        6153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6153)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6153)
  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-m4134452
FEATURES             Location/Qualifiers
     source          1..6153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2500..2520
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2521..2547
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2600..2721
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2611..2630)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2665..2684
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2836..2882
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2912..3048
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3093..3164
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3390..3978)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3470..3489)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4724..4792,4793..5584)
                     /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(4942..4961)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5597..5733
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5885..5956
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5961..6007)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6025..6073
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatggccta cacaagatat
      601 gtggataaca atttttgtgg gccggacgga taccctttag aatgcatcaa agatttgctt
      661 gcaagggcgg gcaaggcttc gtgcactctg tccgagcagc tggacttcat tgataccaag
      721 cgcggtgttt attgttgccg agagcatgaa cacgagatag cctggtatac ggaacggtca
      781 gagaaaagtt acgaactcca aaccccattc gagatcaagc tagctaaaaa gtttgacact
      841 ttcaacggcg aatgtcccaa ttttgtcttt cccctgaact ctattatcaa aacaatacag
      901 cctcgggtgg agaagaagaa attagacgga ttcatggggc gtattaggag cgtataccca
      961 gtggccagcc cgaatgaatg caaccagatg tgtctttcca cactgatgaa gtgcgatcac
     1021 tgtggcgaga cttcttggca aaccggtgac ttcgtaaaag cgacatgcga gttttgtggt
     1081 accgagaatc tgactaagga gggagctacc acatgcggct atttgccaca gaatgcagtg
     1141 gtcaagatct actgtcctgc atgccataac tctgaagtcg gccccgagca ctcgctcgcc
     1201 gaatatcaca atgagagcgg gctaaaaact atactgagga aaggaggtcg caccatcgcc
     1261 tttgggggct gcgttttctc ctacgtgggg tgtcataaca aatgtgccta ttgggttccc
     1321 agagcctccg ctaacattgg gtgcaatcat acgggagtgg tcggggaagg ctcagaaggc
     1381 ttgaacgata acctccttga aattctgcaa aaggagaagg tgaatatcaa tatcgttggc
     1441 gacttcaagt taaatgagga aatcgcgatt attctggcaa gtttttctgc ctcaactagc
     1501 gcctttgtag agacagtgaa aggactcgat tataaagctt tcaaacagat agtcgagagt
     1561 tgtggtaact tcaaggttac aaagggcaag gctaaaaagg gtgcttggaa cattggagag
     1621 cagaagtcaa tcttgagtcc actgtacgcg tttgcatctg aggccgcccg tgtggtcagg
     1681 agcatattca gtcgaaccct agaaaccgca cagaatagcg tgcgggtatt acagaaggcg
     1741 gctattacta ttctcgacgg tatctcgcag tattctctga gactgatcga cgccatgatg
     1801 tttacaagcg atctggcaac gaataactta gtggtcatgg cgtacataac cggaggggtc
     1861 gtgcaactta cctcccagtg gctaacgaac atctttggga cggtttacga gaagctaaaa
     1921 cctgtgctgg actggctcga agagaaattc aaagaaggcg tggaattttt gcgtgacggg
     1981 tgggagatcg taaagttcat atcaacatgt gcttgcgaaa tcgttggcgg acaaatcgta
     2041 acttgcgcaa aagagattaa agaatcggtg cagaccttct ttaaacttgt taacaaattc
     2101 ctggctttgt gtgccgattc tattatcatc gggggagcaa agcttaaagc cctaaattta
     2161 ggcgagacat ttgtgacgca cagtaagggc ctgtatagaa agtgcgtgaa gagtcgcgaa
     2221 gaaaccggac tgctcatgcc cttaaaagcg ccaaaagaaa taattttcct ggaaggggag
     2281 acattgccca ctgaagtgct cactgaggaa gtagttctga aaactgggga tctgcaacct
     2341 ttagagcagc cgacctccga ggccgtcgag gcacctctcg tgggaacccc ggtctgtatt
     2401 aatgggttga tgctgctcga gattaaggat accgaaaaat actgcgcact agcccccaac
     2461 atgatggtga ctaacaacac atttaccctc aagggagggg aaaaccttta ctttcagtcg
     2521 tacccctatg acgtaccaga ttatgcgtaa gctagcttga ctgactgaga tacagcgtac
     2581 cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     2641 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     2701 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     2761 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca
     2821 tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     2881 tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc
     2941 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     3001 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta
     3061 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac
     3121 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct
     3181 tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg
     3241 ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc
     3301 cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg
     3361 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3421 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3481 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3541 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3601 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3661 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3721 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3781 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3841 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3901 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3961 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4021 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4081 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4141 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4201 tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa
     4261 acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg
     4321 ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc
     4381 gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta
     4441 tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa cgatcctctc
     4501 agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg
     4561 ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga
     4621 tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt
     4681 tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt
     4741 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4801 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga
     4861 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat
     4921 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4981 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt
     5041 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5101 catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct
     5161 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5221 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5281 aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat
     5341 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5401 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5461 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5521 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5581 gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     5641 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     5701 tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct
     5761 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
     5821 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
     5881 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
     5941 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     6001 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
     6061 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
     6121 gtcattagtt catagcccat atatggagtt ccg
//
