LOCUS       sequence-297383-        4524 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 4524)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4524)
  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-m4133819
FEATURES             Location/Qualifiers
     source          1..4524
                     /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             275..340
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    393..514
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(404..423)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     458..477
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      629..675
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      705..841
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    886..957
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1183..1771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1263..1282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2517..2585,2586..3377)
                     /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(2735..2754)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3390..3526
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3678..3749
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3754..3800)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3818..3866
                     /note="synthetic polyadenylation signal"
     enhancer        3947..4250
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4252..4454
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4405..4425
                     /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 gatggattat aaggaccacg atggagacta
      301 caaagatcat gacattgatt acaaggacga cgatgacaaa taagctagct tgactgactg
      361 agatacagcg taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac
      421 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
      481 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
      541 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
      601 tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc
      661 ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa
      721 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
      781 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
      841 aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt
      901 cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc
      961 tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag
     1021 atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc
     1081 ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca
     1141 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1201 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1261 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1321 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1381 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1441 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1501 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1561 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1621 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1681 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1741 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1801 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1861 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1921 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1981 ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc
     2041 ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc
     2101 gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta
     2161 ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct
     2221 caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga
     2281 gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc
     2341 ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac
     2401 ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc
     2461 gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga
     2521 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2581 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag
     2641 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2701 aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2761 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2821 agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2881 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgattggag
     2941 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3001 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3061 tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc
     3121 ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3181 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3241 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3301 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3361 tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga
     3421 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     3481 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt
     3541 taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg
     3601 cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca
     3661 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt
     3721 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga
     3781 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc
     3841 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt
     3901 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     3961 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     4021 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     4081 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     4141 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     4201 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt
     4261 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     4321 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     4381 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     4441 atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca
     4501 ctcgacacac ccgccagcgg ccgc
//
