LOCUS       sequence-300693-        7197 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 7197)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7197)
  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-m4252920
FEATURES             Location/Qualifiers
     source          1..7197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        97..400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    481..529
                     /note="synthetic polyadenylation signal"
     terminator      547..593
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    598..669
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(821..957)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(970..1761,1762..1830))
                     /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     1593..1612
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    2096..2144
                     /note="synthetic polyadenylation signal"
     CDS             complement(2171..3199)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(3203..4412)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(3347..3366)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4912..5500
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5401..5420
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5726..5797
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5842..5978)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(6008..6054)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6169..6290
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6206..6225)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6260..6279
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(6383..6400)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(6749..6769)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(join(7090..7197,1..95))
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(7119..7139)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta
       61 tccacgccca ttgatgtact gccaaaaccg catcagcatg gtaatagcga tgactaatac
      121 gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc
      181 gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg
      241 tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc
      301 tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg
      361 cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct
      421 atgaactaat gaccccgtaa ttgattacta gatctggcta cactcgatca gcagttagat
      481 aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgcaac
      541 gattgatagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgaattgag
      601 aggtagctga agctgctgtt atgaccacta gagggaagaa gatacctgtg gagctaatgg
      661 tccaagatgt cgggagccgc gatcgctgcc aaaaaggtaa gctgacgata gctgacgaga
      721 ttcgatgccc gacgagcaaa tgagcgccac ggctgttcgc gtaagctcga aacgacggag
      781 attcgataga gccagagtcg agtttaaacg attcgggatc ttaaggataa atttctggta
      841 aggaggacac gtatggaagt gggcaagttg gggaagccgt atccgttgct gaatctggca
      901 tatgtgggag tataagacgc gcagcgtcgc atcaggcatt tttttctgcg ccaatgcacc
      961 tagaatcggt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1021 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     1081 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     1141 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     1201 ctccatccag tctatcaact gttgccggga agctagagta agtagttcgc cagttaatag
     1261 tttacgcaag gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     1321 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     1381 caaaaaagcg gttagctcct tcggtcctcc aatcgttgtc agaagtaagt tggccgcagt
     1441 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     1501 atgcttttct gtgactggtg aatactcaac caagtcattc tgagaatagt gtatgcggcg
     1561 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     1621 aaaagtgctc atcattggaa agcgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     1681 gttgagatcc agttcgatgt aacccactcg cgcacccaac tgatcttcag catcttttac
     1741 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     1801 aagggcgaca cggaaatgtt gaatactcat gcgaaagcct cctgctgatc ctgtctcttg
     1861 atagatgttt gcaaaagcta ggactcgatt atacgttgac cgatcagact atcaatatct
     1921 aggtttaaac agatcggtac gctgccgtga ccaggcttga gtacaatatc tgacgattgg
     1981 acttcctggg tcccaagctg gattccgact ggctgactgc caagcaacga tatggatcgt
     2041 cgagactcgc actgagagga tcgttctcgg tacaccaggc gcgcctagcg ttacacacac
     2101 aaaaaaccaa cacacatcca tcttcgatgg atagcgattt tattatctaa gactaggctt
     2161 acatactagc ttattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg
     2221 cgagtacttc tacacagcca tcggtccaga cggcggcgct tctgcgggcg atttgtgtac
     2281 gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg
     2341 catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca
     2401 tatacgcccg gagccggggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg
     2461 tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt
     2521 gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg
     2581 tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct
     2641 cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca
     2701 tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg
     2761 tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat
     2821 cggcggcagc gatggcatcc attgcctccg cgaccggctg cagaacagcg ggcagttcgg
     2881 tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc
     2941 tctcgctgaa ctccccaatg tcaagcactt ccggaatcgg gagcgcggcg gatgcaaagt
     3001 gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat
     3061 atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca
     3121 tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga
     3181 gttcaggctt tttacccatg ttgtctaaca aaaaagccaa aaacggccag aatttagcgg
     3241 acaatttaca agtctaacac tgaaaattac atattgaccc aaatgattac atttcaaaag
     3301 gtgcctaaaa aacttcacaa aacacactcg ccaaccccga gcgcatagtt caaaaccgga
     3361 gcttcagcta cttaagaaga taggtacata aaaccgacca aagaaactga cgcctcactt
     3421 atccctcccc tcaccagagg tccggcgcct gtcgattcag gagagcctac cctaggcccg
     3481 aaccctgcgt cctgcgacgg agaaaagcct accgcacacc taccggctgg tggccccacc
     3541 ctgcattata agccaacaga acgggtgacg tcacgacacg acgagggcgc gcgctcccaa
     3601 aggtacgggt gcactgccca acggcaccgc cataactgcc gcccccgcaa cagacgacaa
     3661 accgagttct ccagtcagtg acaaacttca cgtcagggtc cccagatggt gcccagccca
     3721 tctcacccga ataagagctt tcccgcatta gcgaagggct caagaccttg ggttcttgcc
     3781 gcccaccatg ccccccacct tgtttcaacg acctcacagc ccgcctcaca agcgtcttcc
     3841 attcaagact cgggaacagc cgccattttg ctgcgctccc cccaaccccc agttcagggc
     3901 aaccttgctc gcggacccag actacagccc ttggcggtct ctccacacgc ttccgtccca
     3961 ccgagcggcc cggcggccac gaaagccccg gccagcccag cagcccgcta ctcaccaagt
     4021 gacgatcaca gcgatccaca aacaagaacc gcgacccaaa tcccggctgc gacggaacta
     4081 gctgtgccac acccggcgcg tccttatata atcatcggcg ttcaccgccc cacggagatc
     4141 cctccgcaga atcgccgaga agggactact tttcctcgcc tgttccgctc tctggaaaga
     4201 aaaccagtgc cctagagtca cccaagtccc gtcctaaaat gtccttctgc tgatactggg
     4261 gttctaaggc cgagtcttat gagcagcggg ccgctgtcct gagcgtccgg gcggaaggat
     4321 caggacgctc ctgcgccctt cgtctgacgt ggcagcgctc gccgtgacga ggggggcgcc
     4381 cgcgggaggc gccaaaaccc ggcgcggagg ccaggcgcgc cccgatttcg atattcctcg
     4441 gatctcggat agatcgggat tgaggatgta ggttgaactt ctctcgggtg atcccgatct
     4501 cggatattcg ggattggagt aatacctggc gataggcgct gccaagcaat agccaaggcg
     4561 caaggccggc caagagactt gcgatagttc gttcgagagg tagcctgcaa gatggatcgg
     4621 acgaaaggtt tgaagatttt ccgacgaggg ccttggagag ttcggaaatt taaatgcaac
     4681 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4741 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4801 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4861 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4921 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4981 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5041 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5101 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5161 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5221 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5281 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5341 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5401 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5461 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5521 tttacggttc ctggcctttt gctggccttt tgctcacatg atttaaatga aactcgtaat
     5581 cggagaacgg gtgcaatagc aaggaccaga tcgaacttga ctgacgcaat tctgagcgta
     5641 gtcgaaaccc agacaagtcg atctcgccaa gtagctgact tgactggtta attaagtttc
     5701 gatagcccaa ggtaaccaat gagattaggg agggaagaaa gtttgatgaa ctacttctga
     5761 cccctagtgg tgtccagaaa agaccattaa aggaatgtct tgatcgaaat cgagagatcg
     5821 cctgcaggat agctgacgac cttaaggata aatttctggt aaggaggaca cgtatggaag
     5881 tgggcaagtt ggggaagccg tatccgttgc tgaatctggc atatgtggga gtataagacg
     5941 cgcagcgtcg catcaggcat ttttttctgc gccaatgccg gtagatagca atccggcccg
     6001 aggggcacaa aaaacccctc aagacccgtt tagaggcccc aaggggttat gctacgatac
     6061 cgatagagat gggccaatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     6121 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     6181 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6241 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     6301 gagctgaagg tacgctgtat ctcagtcagt caagctagca gaggcctgcg gatccttagc
     6361 cgcgtggcac gagcccgctg ctatggtgat ggtgatggtg catgtgcccc gagcctgaac
     6421 ctgccaggtt agcgtccaag aactctttta gctggccctt agacagggct cccacttttg
     6481 tcgcggcaac ttccccattc ttaaagagca ggagggtagg tattcccctg atgccgtatt
     6541 taggtgccgt ccctgggttc tgatctatat taagcttagc gacagttaat tttccttgat
     6601 actcgtcggc gatctcgtcc agaatgggtg cgatcatctt gcacgggcca caccattcag
     6661 cccagaaatc taccagtatc gcaccatcgg ccttcaacac gtctgtatca aaggagtcat
     6721 cggttagatg aatgattttg tcactcatcg actgaaagta aaggttttct aaggttgccc
     6781 tgttgtccag catttcttca caaagcttgt ttatgtctac agcaccctgc atggaaagca
     6841 aaacagaaag tagtgaaacc attttttcaa aggcttcagt agtatcttta gctaagagaa
     6901 tgtcattgtg taactggaca cattgagccc acaatttaga tgatgattct actctgagtt
     6961 gttgcaaaac tgagagtaag actactgatg tgcactttac atctgacatt ttcatggtgg
     7021 cggccgctgg cgggtgtgtc gagtggatgg taggatcgac aaagatctga tctgacggtt
     7081 cactaaacca gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     7141 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaa
//
