LOCUS       sequence-301676-        7368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7368)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7368)
  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-m4252946
FEATURES             Location/Qualifiers
     source          1..7368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    72..120
                     /note="synthetic polyadenylation signal"
     terminator      138..184
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    189..260
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(412..548)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(561..1352,1353..1421))
                     /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     1184..1203
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1687..1735
                     /note="synthetic polyadenylation signal"
     CDS             complement(1762..2790)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2794..4003)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2938..2957)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4503..5091
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4992..5011
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5317..5388
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5433..5569)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5599..5645)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5760..5881
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5797..5816)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5851..5870
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(6346..6366)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(6394..6471)
                     /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             complement(6505..6525)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(6852..7054)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6881..6901)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7056..7359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatatgggc tatgaactaa tgaccccgta attgattact agatctggct acactcgatc
       61 agcagttaga taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg
      121 tgtaacgcaa cgattgatag cataacccct tggggcctct aaacgggtct tgaggggttt
      181 tttgaattga gaggtagctg aagctgctgt tatgaccact agagggaaga agatacctgt
      241 ggagctaatg gtccaagatg tcgggagccg cgatcgctgc caaaaaggta agctgacgat
      301 agctgacgag attcgatgcc cgacgagcaa atgagcgcca cggctgttcg cgtaagctcg
      361 aaacgacgga gattcgatag agccagagtc gagtttaaac gattcgggat cttaaggata
      421 aatttctggt aaggaggaca cgtatggaag tgggcaagtt ggggaagccg tatccgttgc
      481 tgaatctggc atatgtggga gtataagacg cgcagcgtcg catcaggcat ttttttctgc
      541 gccaatgcac ctagaatcgg ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      601 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
      661 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
      721 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
      781 actttatccg cctccatcca gtctatcaac tgttgccggg aagctagagt aagtagttcg
      841 ccagttaata gtttacgcaa ggttgttgcc attgctacag gcatcgtggt gtcacgctcg
      901 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
      961 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc caatcgttgt cagaagtaag
     1021 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1081 ccatccgtaa gatgcttttc tgtgactggt gaatactcaa ccaagtcatt ctgagaatag
     1141 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1201 agcagaactt taaaagtgct catcattgga aagcgttctt cggggcgaaa actctcaagg
     1261 atcttaccgc tgttgagatc cagttcgatg taacccactc gcgcacccaa ctgatcttca
     1321 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1381 aaaaagggaa taagggcgac acggaaatgt tgaatactca tgcgaaagcc tcctgctgat
     1441 cctgtctctt gatagatgtt tgcaaaagct aggactcgat tatacgttga ccgatcagac
     1501 tatcaatatc taggtttaaa cagatcggta cgctgccgtg accaggcttg agtacaatat
     1561 ctgacgattg gacttcctgg gtcccaagct ggattccgac tggctgactg ccaagcaacg
     1621 atatggatcg tcgagactcg cactgagagg atcgttctcg gtacaccagg cgcgcctagc
     1681 gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta
     1741 agactaggct tacatactag cttattcctt tgccctcgga cgagtgctgg ggcgtcggtt
     1801 tccactatcg gcgagtactt ctacacagcc atcggtccag acggcggcgc ttctgcgggc
     1861 gatttgtgta cgcccgacag tcccggctcc ggatcggacg attgcgtcgc atcgaccctg
     1921 cgcccaagct gcatcatcga aattgccgtc aaccaagctc tgatagagtt ggtcaagacc
     1981 aatgcggagc atatacgccc ggagccgggg cgatcctgca agctccggat gcctccgctc
     2041 gaagtagcgc gtctgctgct ccatacaagc caaccacggc ctccagaaga agatgttggc
     2101 gacctcgtat tgggaatccc cgaacatcgc ctcgctccag tcaatgaccg ctgttatgcg
     2161 gccattgtcc gtcaggacat tgttggagcc gaaatccgcg tgcacgaggt gccggacttc
     2221 ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc tgcgcgacgg acgcactgac
     2281 ggtgtcgtcc atcacagttt gccagtgata cacatgggga tcagcaatcg cgcatatgaa
     2341 atcacgccat gtagtgtatt gaccgattcc ttgcggtccg aatgggccga acccgctcgt
     2401 ctggctaaga tcggcggcag cgatggcatc cattgcctcc gcgaccggct gcagaacagc
     2461 gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc tgtgcacggc gggagatgca
     2521 ataggtcagg ctctcgctga actccccaat gtcaagcact tccggaatcg ggagcgcggc
     2581 ggatgcaaag tgccgataaa cataacgatc tttgtagaaa ccatcggcgc agctatttac
     2641 ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa gcacgagatt cttcgccctc
     2701 cgagagctgc atcaggtcgg agacgctgtc gaacttttcg atcagaaact tctcgacaga
     2761 cgtcgcggtg agttcaggct ttttacccat gttgtctaac aaaaaagcca aaaacggcca
     2821 gaatttagcg gacaatttac aagtctaaca ctgaaaatta catattgacc caaatgatta
     2881 catttcaaaa ggtgcctaaa aaacttcaca aaacacactc gccaaccccg agcgcatagt
     2941 tcaaaaccgg agcttcagct acttaagaag ataggtacat aaaaccgacc aaagaaactg
     3001 acgcctcact tatccctccc ctcaccagag gtccggcgcc tgtcgattca ggagagccta
     3061 ccctaggccc gaaccctgcg tcctgcgacg gagaaaagcc taccgcacac ctaccggctg
     3121 gtggccccac cctgcattat aagccaacag aacgggtgac gtcacgacac gacgagggcg
     3181 cgcgctccca aaggtacggg tgcactgccc aacggcaccg ccataactgc cgcccccgca
     3241 acagacgaca aaccgagttc tccagtcagt gacaaacttc acgtcagggt ccccagatgg
     3301 tgcccagccc atctcacccg aataagagct ttcccgcatt agcgaagggc tcaagacctt
     3361 gggttcttgc cgcccaccat gccccccacc ttgtttcaac gacctcacag cccgcctcac
     3421 aagcgtcttc cattcaagac tcgggaacag ccgccatttt gctgcgctcc ccccaacccc
     3481 cagttcaggg caaccttgct cgcggaccca gactacagcc cttggcggtc tctccacacg
     3541 cttccgtccc accgagcggc ccggcggcca cgaaagcccc ggccagccca gcagcccgct
     3601 actcaccaag tgacgatcac agcgatccac aaacaagaac cgcgacccaa atcccggctg
     3661 cgacggaact agctgtgcca cacccggcgc gtccttatat aatcatcggc gttcaccgcc
     3721 ccacggagat ccctccgcag aatcgccgag aagggactac ttttcctcgc ctgttccgct
     3781 ctctggaaag aaaaccagtg ccctagagtc acccaagtcc cgtcctaaaa tgtccttctg
     3841 ctgatactgg ggttctaagg ccgagtctta tgagcagcgg gccgctgtcc tgagcgtccg
     3901 ggcggaagga tcaggacgct cctgcgccct tcgtctgacg tggcagcgct cgccgtgacg
     3961 aggggggcgc ccgcgggagg cgccaaaacc cggcgcggag gccaggcgcg ccccgatttc
     4021 gatattcctc ggatctcgga tagatcggga ttgaggatgt aggttgaact tctctcgggt
     4081 gatcccgatc tcggatattc gggattggag taatacctgg cgataggcgc tgccaagcaa
     4141 tagccaaggc gcaaggccgg ccaagagact tgcgatagtt cgttcgagag gtagcctgca
     4201 agatggatcg gacgaaaggt ttgaagattt tccgacgagg gccttggaga gttcggaaat
     4261 ttaaatgcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4321 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4381 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4441 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4501 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4561 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4621 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4681 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4741 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4801 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4861 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4921 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4981 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5041 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5101 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gatttaaatg
     5161 aaactcgtaa tcggagaacg ggtgcaatag caaggaccag atcgaacttg actgacgcaa
     5221 ttctgagcgt agtcgaaacc cagacaagtc gatctcgcca agtagctgac ttgactggtt
     5281 aattaagttt cgatagccca aggtaaccaa tgagattagg gagggaagaa agtttgatga
     5341 actacttctg acccctagtg gtgtccagaa aagaccatta aaggaatgtc ttgatcgaaa
     5401 tcgagagatc gcctgcagga tagctgacga ccttaaggat aaatttctgg taaggaggac
     5461 acgtatggaa gtgggcaagt tggggaagcc gtatccgttg ctgaatctgg catatgtggg
     5521 agtataagac gcgcagcgtc gcatcaggca tttttttctg cgccaatgcc ggtagatagc
     5581 aatccggccc gaggggcaca aaaaacccct caagacccgt ttagaggccc caaggggtta
     5641 tgctacgata ccgatagaga tgggccaata ccacatttgt agaggtttta cttgctttaa
     5701 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     5761 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5821 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5881 atcatgtctg tgagctgaag gtacgctgta tctcagtcag tcaagctagc agaggcctgc
     5941 ggatccttat tgatggttcg cgtctacttt cggggcctgg gcatcattga gctttttggc
     6001 ttccgcaagc aggttggcac tctgtgatgg atcgtctttt agggactgga taaatgcatt
     6061 tctttgttcc tcattcagat taggaaggtg caatatctcg tagaaagcat tctgctgctc
     6121 cttgttaaac ttgttatcaa cttttggagc ctgtgcgtca ttcagcttct tggcttcggc
     6181 caggagattt gcagactgcg agggatcgtc tttcaggctc tgaatgaaag cgttgcgttg
     6241 ctcttcgttc agatttggta agtgaaggat ctcatagaaa gcgttctgct gttctttatt
     6301 aaacttgtta tcgacggcct cgtcatgttg tgccaagcca aattcaccct ggaaatagag
     6361 attctcggag gcggtcgtag gaatgtcgta atctagggcg ccgcttgaac tgatcttttt
     6421 gaaccggtta gccgcagaca ctgcaataaa gttcttcttc catctccttt tccagggatt
     6481 tatcagcccc gggattcgcc gtcccgactg aaagtaaagg ttttcttgca gggtggctcg
     6541 gttgtctaac atctcttcgc aaagtttgtt gatgtctact gctccttgca tgctcagtag
     6601 aacagatagc aagctgacca tcttctcgaa agcttccgta gtatctttcg cgagcaatat
     6661 atcattgtgc agctgcacgc actgggccca gagctttgac gaactctcca cccgcagctg
     6721 ttgaagcact gataacagaa ctacggatgt acacttgacg tcagacattt tgctcatggt
     6781 ggcggccgct ggcgggtgtg tcgagtggat ggtaggatcg acaaagatct gatctgacgg
     6841 ttcactaaac cagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc
     6901 gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac
     6961 aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat
     7021 ccacgcccat tgatgtactg ccaaaaccgc atcagcatgg taatagcgat gactaatacg
     7081 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
     7141 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
     7201 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     7261 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     7321 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactcc
//
