LOCUS       sequence-295133-        4497 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 4497)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4497)
  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-m4134316
FEATURES             Location/Qualifiers
     source          1..4497
                     /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..313
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    366..487
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(377..396)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     431..450
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      602..648
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      678..814
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    859..930
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1156..1744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1236..1255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2490..2558,2559..3350)
                     /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(2708..2727)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3363..3499
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3651..3722
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3727..3773)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3791..3839
                     /note="synthetic polyadenylation signal"
     enhancer        3920..4223
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4225..4427
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4378..4398
                     /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 ggggaaacct atacccaatc cacttctcgg
      301 cttagattct acctaagcta gcttgactga ctgagataca gcgtaccttc agctcacaga
      361 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
      421 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
      481 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
      541 ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc
      601 gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc
      661 cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
      721 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
      781 atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc
      841 tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga
      901 agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac
      961 ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc
     1021 tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac
     1081 gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1141 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1201 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1261 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1321 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1381 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1441 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1501 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1561 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1621 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1681 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1741 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1801 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1861 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1921 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1981 atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat
     2041 ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct
     2101 tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc
     2161 gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg
     2221 aatatcgaaa tcggggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     2281 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     2341 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     2401 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     2461 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     2521 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2581 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     2641 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     2701 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2761 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     2821 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2881 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     2941 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3001 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     3061 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     3121 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3181 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3241 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3301 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     3361 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     3421 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     3481 accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc
     3541 tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat
     3601 ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac
     3661 cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc
     3721 tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg
     3781 ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta
     3841 tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata
     3901 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     3961 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     4021 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     4081 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     4141 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     4201 tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat
     4261 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     4321 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     4381 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     4441 gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
