LOCUS       sequence-295819-        4482 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 4482)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4482)
  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-m4133422
FEATURES             Location/Qualifiers
     source          1..4482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             251..271
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             272..298
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    351..472
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(362..381)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     416..435
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      587..633
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      663..799
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    844..915
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1141..1729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1221..1240)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2475..2543,2544..3335)
                     /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(2693..2712)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3348..3484
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3636..3707
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3712..3758)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3776..3824
                     /note="synthetic polyadenylation signal"
     enhancer        3905..4208
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4210..4412
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4363..4383
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaaa atgtcagatg taaagtgcac atcagtagtc ttactctcag ttttgcaaca
       61 actcagagta gaatcatcat ctaaattgtg ggctcaatgt gtccagttac acaatgacat
      121 tctcttagct aaagatacta ctgaagcctt tgaaaaaatg gtttcactac tttctgtttt
      181 gctttccatg cagggtgctg tagacataaa caagctttgt gaagaaatgc tggacaacag
      241 ggcaacctta gaaaaccttt actttcagtc gtacccctat gacgtaccag attatgcgta
      301 agctagcttg actgactgag atacagcgta ccttcagctc acagacatga taagatacat
      361 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
      421 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
      481 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
      541 gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt
      601 ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac
      661 cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
      721 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
      781 accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca
      841 agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac
      901 tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca
      961 agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag
     1021 tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat
     1081 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1141 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1201 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1261 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1321 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1381 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1441 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1501 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1561 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     1621 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1681 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1741 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     1801 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     1861 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     1921 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa
     1981 ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta
     2041 cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc
     2101 agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag
     2161 aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg
     2221 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     2281 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     2341 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     2401 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     2461 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2521 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2581 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2641 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2701 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2761 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     2821 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2881 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2941 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3001 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     3061 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     3121 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3181 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3241 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3301 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     3361 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     3421 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     3481 ttaagatccc gaatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc
     3541 ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt
     3601 cagcttacct ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg
     3661 tatcttcttc cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc
     3721 cctcaagacc cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac
     3781 aaaaaaccaa cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg
     3841 agtgtagcca gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     3901 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     3961 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     4021 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     4081 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     4141 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     4201 ttaccatgct gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     4261 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     4321 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     4381 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt
     4441 tgtcgatcct accatccact cgacacaccc gccagcggcc gc
//
