LOCUS       sequence-317258-        6162 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 6162)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6162)
  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-m4134051
FEATURES             Location/Qualifiers
     source          1..6162
                     /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"
     CDS             2494..2514
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2515..2556
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    2609..2730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2620..2639)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2674..2693
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2845..2891
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2921..3057
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3102..3173
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3399..3987)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3479..3498)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4733..4801,4802..5593)
                     /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(4951..4970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5606..5742
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5894..5965
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5970..6016)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6034..6082
                     /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 ccatgtacac tcgctatgtc
      601 gataacaact tctgtggccc tgatggctac cctcttgagt gcattaaaga ccttctagca
      661 cgtgctggta aagcttcatg cactttgtcc gaacaactgg actttattga cactaagagg
      721 ggtgtatact gctgccgtga acatgagcat gaaattgctt ggtacacgga acgttctgaa
      781 aagagctatg aattgcagac accttttgaa attaaattgg caaagaaatt tgacaccttc
      841 aatggggaat gtccaaattt tgtatttccc ttaaattcca taatcaagac tattcaacca
      901 agggttgaaa agaaaaagct tgatggcttt atgggtagaa ttcgatctgt ctatccagtt
      961 gcgtcaccaa atgaatgcaa ccaaatgtgc ctttcaactc tcatgaagtg tgatcattgt
     1021 ggtgaaactt catggcagac gggcgatttt gttaaagcca cttgcgaatt ttgtggcact
     1081 gagaatttga ctaaagaagg tgccactact tgtggttact taccccaaaa tgctgttgtt
     1141 aaaatttatt gtccagcatg tcacaattca gaagtaggac ctgagcatag tcttgccgaa
     1201 taccataatg aatctggctt gaaaaccatt cttcgtaagg gtggtcgcac tattgccttt
     1261 ggaggctgtg tgttctctta tgttggttgc cataacaagt gtgcctattg ggttccacgt
     1321 gctagcgcta acataggttg taaccataca ggtgttgttg gagaaggttc cgaaggtctt
     1381 aatgacaacc ttcttgaaat actccaaaaa gagaaagtca acatcaatat tgttggtgac
     1441 tttaaactta atgaagagat cgccattatt ttggcatctt tttctgcttc cacaagtgct
     1501 tttgtggaaa ctgtgaaagg tttggattat aaagcattca aacaaattgt tgaatcctgt
     1561 ggtaatttta aagttacaaa aggaaaagct aaaaaaggtg cctggaatat tggtgaacag
     1621 aaatcaatac tgagtcctct ttatgcattt gcatcagagg ctgctcgtgt tgtacgatca
     1681 attttctccc gcactcttga aactgctcaa aattctgtgc gtgttttaca gaaggccgct
     1741 ataacaatac tagatggaat ttcacagtat tcactgagac tcattgatgc tatgatgttc
     1801 acatctgatt tggctactaa caatctagtt gtaatggcct acattacagg tggtgttgtt
     1861 cagttgactt cgcagtggct aactaacatc tttggcactg tttatgaaaa actcaaaccc
     1921 gtccttgatt ggcttgaaga gaagtttaag gaaggtgtag agtttcttag agacggttgg
     1981 gaaattgtta aatttatctc aacctgtgct tgtgaaattg tcggtggaca aattgtcacc
     2041 tgtgcaaagg aaattaagga gagtgttcag acattcttta agcttgtaaa taaatttttg
     2101 gctttgtgtg ctgactctat cattattggt ggagctaaac ttaaagcctt gaatttaggt
     2161 gaaacatttg tcacgcactc aaagggattg tacagaaagt gtgttaaatc cagagaagaa
     2221 actggcctac tcatgcctct aaaagcccca aaagaaatta tcttcttaga gggagaaaca
     2281 cttcccacag aagtgttaac agaggaagtt gtcttgaaaa ctggtgattt acaaccatta
     2341 gaacaaccta ctagtgaagc tgttgaagct ccattggttg gtacaccagt ttgtattaac
     2401 gggcttatgt tgctcgaaat caaagacaca gaaaagtact gtgcccttgc acctaatatg
     2461 atggtaacaa acaatacctt cacactcaaa ggcgaaaacc tttactttca gtcggggaaa
     2521 cctataccca atccacttct cggcttagat tctacctaag ctagcttgac tgactgagat
     2581 acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca
     2641 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2701 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2761 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     2821 attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga
     2881 ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat
     2941 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     3001 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt
     3061 cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt
     3121 tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat
     3181 tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg
     3241 acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc
     3301 tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa
     3361 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3421 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3481 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3541 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3601 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3661 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3721 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3781 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3841 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3901 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3961 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4021 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4081 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4141 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4201 ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa
     4261 aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa
     4321 gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac
     4381 tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat
     4441 cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg taccgagaac
     4501 gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag ccagtcggaa
     4561 tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct ggtcacggca
     4621 gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt ataatcgagt
     4681 cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc gcatgagtat
     4741 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4801 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg
     4861 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4921 ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4981 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5041 ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5101 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga ttggaggacc
     5161 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5221 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5281 aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag cttcccggca
     5341 acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5401 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5461 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5521 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5581 taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct
     5641 gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc
     5701 ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga atcgtttaaa
     5761 ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag ccgtggcgct
     5821 catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga
     5881 tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc
     5941 ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc
     6001 ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc
     6061 ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc
     6121 aattacgggg tcattagttc atagcccata tatggagttc cg
//
