LOCUS       sequence-300595-        4473 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 4473)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4473)
  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-m4133796
FEATURES             Location/Qualifiers
     source          1..4473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             112..132
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             133..150
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    203..324
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(214..233)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     268..287
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      439..485
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      515..651
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    696..767
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(993..1581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1073..1092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2327..2395,2396..3187)
                     /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(2545..2564)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3200..3336
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3488..3559
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3564..3610)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3628..3676
                     /note="synthetic polyadenylation signal"
     enhancer        3757..4060
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4062..4264
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4215..4235
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 actgaagcct ttgaaaaaat ggtttcacta ctttctgttt tgctttccat gcagggtgct
       61 gtagacataa acaagctttg tgaagaaatg ctggacaaca gggcaacctt agaaaacctt
      121 tactttcagt cgcaccatca ccatcaccat taagctagct tgactgactg agatacagcg
      181 taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga
      241 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
      301 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
      361 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc
      421 ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
      481 ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga
      541 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
      601 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag
      661 ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga
      721 caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta
      781 ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt
      841 ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc
      901 acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc
      961 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1021 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1081 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1141 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1201 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1261 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1321 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1381 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1441 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1501 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1561 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1621 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1681 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1741 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1801 cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt
     1861 caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct
     1921 tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat
     1981 cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat
     2041 ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct
     2101 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     2161 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     2221 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     2281 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca
     2341 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2401 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat
     2461 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc
     2521 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     2581 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc
     2641 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2701 aaccatgagt gataacactg cggccaactt acttctgaca acgattggag gaccgaagga
     2761 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2821 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     2881 aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt
     2941 gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3001 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3061 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3121 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3181 ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     3241 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     3301 ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt taaactcgac
     3361 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     3421 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     3481 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     3541 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     3601 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     3661 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     3721 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     3781 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     3841 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     3901 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     3961 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     4021 ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt tttggcagta
     4081 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     4141 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     4201 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     4261 agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca ctcgacacac
     4321 ccgccagcgg ccgccaccat gaaaatgtca gatgtaaagt gcacatcagt agtcttactc
     4381 tcagttttgc aacaactcag agtagaatca tcatctaaat tgtgggctca atgtgtccag
     4441 ttacacaatg acattctctt agctaaagat act
//
