LOCUS       sequence-313252-        9033 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 9033)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9033)
  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-m4134411
FEATURES             Location/Qualifiers
     source          1..9033
                     /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             2554..2631
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             2659..2679
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    3144..3265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3155..3174)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3209..3228
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3380..3426
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3456..3592
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3637..3708
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3934..4522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4014..4033)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5022..6231
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6068..6087
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             6235..7263
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    7290..7338
                     /note="synthetic polyadenylation signal"
     CDS             join(7604..7672,7673..8464)
                     /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(7822..7841)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8477..8613
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8765..8836
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8841..8887)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8905..8953
                     /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 ggacggcgaa tcccggggct gataaatccc tggaaaagga gatggaagaa gaactttatt
     2581 gcagtgtctg cggctaaccg gttcaaaaag atcagttcaa gcggcgccct agattacgac
     2641 attcctacga ccgcctccga gaatctctat ttccagggtg aatttggctt ggcacaacat
     2701 gacgaggccg tcgataacaa gtttaataaa gaacagcaga acgctttcta tgagatcctt
     2761 cacttaccaa atctgaacga agagcaacgc aacgctttca ttcagagcct gaaagacgat
     2821 ccctcgcagt ctgcaaatct cctggccgaa gccaagaagc tgaatgacgc acaggctcca
     2881 aaagttgata acaagtttaa caaggagcag cagaatgctt tctacgagat attgcacctt
     2941 cctaatctga atgaggaaca aagaaatgca tttatccagt ccctaaaaga cgatccatca
     3001 cagagtgcca acctgcttgc ggaagccaaa aagctcaatg atgcccaggc cccgaaagta
     3061 gacgcgaacc atcaataagg atccgcaggc ctctgctagc ttgactgact gagatacagc
     3121 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3181 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3241 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3301 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     3361 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     3421 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     3481 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     3541 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     3601 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     3661 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     3721 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     3781 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     3841 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     3901 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3961 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4021 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4081 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4141 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4201 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4261 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4321 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4381 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4441 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4501 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4561 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4621 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4681 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4741 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     4801 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     4861 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     4921 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     4981 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt
     5041 ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg
     5101 caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac
     5161 tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct
     5221 agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc
     5281 gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg
     5341 ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat
     5401 cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg
     5461 ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc
     5521 cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc
     5581 ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg
     5641 gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg ggaaagctct
     5701 tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac
     5761 tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag
     5821 tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg
     5881 gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc
     5941 aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg
     6001 tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt
     6061 aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa
     6121 gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag
     6181 acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga caacatgggt
     6241 aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc
     6301 gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta
     6361 ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt
     6421 tatgtttatc ggcactttgc atccgccgcg ctcccgattc cggaagtgct tgacattggg
     6481 gagttcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa
     6541 gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc aatggatgcc
     6601 atcgctgccg ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc
     6661 ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac
     6721 tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg
     6781 atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc
     6841 aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg
     6901 ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt
     6961 atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccccgg
     7021 ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc
     7081 aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc
     7141 gggactgtcg ggcgtacaca aatcgcccgc agaagcgccg ccgtctggac cgatggctgt
     7201 gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa
     7261 taagctagta tgtaagccta gtcttagata ataaaatcgc tatccatcga agatggatgt
     7321 gtgttggttt tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa cgatcctctc
     7381 agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg
     7441 ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga
     7501 tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt
     7561 tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt
     7621 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7681 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga
     7741 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat
     7801 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     7861 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt
     7921 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     7981 catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct
     8041 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     8101 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     8161 aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat
     8221 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     8281 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     8341 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     8401 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     8461 gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     8521 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     8581 tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct
     8641 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
     8701 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
     8761 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
     8821 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     8881 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
     8941 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
     9001 gtcattagtt catagcccat atatggagtt ccg
//
