LOCUS       sequence-310621-        6189 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 6189)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6189)
  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-m4134143
FEATURES             Location/Qualifiers
     source          1..6189
                     /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             2518..2583
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2636..2757
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2647..2666)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2701..2720
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2872..2918
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2948..3084
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3129..3200
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3426..4014)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3506..3525)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4760..4828,4829..5620)
                     /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(4978..4997)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5633..5769
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5921..5992
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5997..6043)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6061..6109
                     /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 gtcgatggat
     2521 tataaggacc acgatggaga ctacaaagat catgacattg attacaagga cgacgatgac
     2581 aaataagcta gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga
     2641 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     2701 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     2761 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     2821 agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac
     2881 cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta
     2941 tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     3001 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     3061 tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc
     3121 gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat
     3181 caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc
     3241 agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc
     3301 gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt
     3361 taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3421 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3481 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3541 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3601 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3661 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3721 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3781 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3841 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3901 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3961 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4021 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4081 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4141 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4201 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt
     4261 ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca
     4321 ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc
     4381 ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc
     4441 gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa
     4501 tcggggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     4561 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     4621 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     4681 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     4741 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     4801 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4861 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4921 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     4981 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5041 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     5101 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5161 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5221 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5281 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     5341 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     5401 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5461 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5521 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5581 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     5641 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     5701 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     5761 tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     5821 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     5881 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     5941 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     6001 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     6061 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     6121 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     6181 ggagttccg
//
