LOCUS       sequence-300699-        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             join(740..808,809..1600)
                     /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(958..977)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      1613..1749
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1901..1972
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1977..2023)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    2041..2089
                     /note="synthetic polyadenylation signal"
     enhancer        2170..2473
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2475..2677
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2628..2648
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2998..3018
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3022..3087
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    3140..3261
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3151..3170)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3205..3224
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3376..3422
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3452..3588
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3633..3704
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3930..4518)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4010..4029)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
       61 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
      121 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
      181 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt
      241 ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca
      301 ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc
      361 ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc
      421 gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa
      481 tcggggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
      541 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
      601 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
      661 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
      721 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
      781 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
      841 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
      901 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
      961 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     1021 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     1081 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1141 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     1201 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     1261 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     1321 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     1381 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     1441 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     1501 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1561 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     1621 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     1681 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     1741 tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     1801 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     1861 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     1921 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     1981 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     2041 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     2101 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     2161 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     2221 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     2281 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     2341 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     2401 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     2461 cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     2521 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     2581 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     2641 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag
     2701 atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgccac catgaaaatg
     2761 tcagatgtaa agtgcacatc agtagtctta ctctcagttt tgcaacaact cagagtagaa
     2821 tcatcatcta aattgtgggc tcaatgtgtc cagttacaca atgacattct cttagctaaa
     2881 gatactactg aagcctttga aaaaatggtt tcactacttt ctgttttgct ttccatgcag
     2941 ggtgctgtag acataaacaa gctttgtgaa gaaatgctgg acaacagggc aaccttagaa
     3001 aacctttact ttcagtcgat ggattataag gaccacgatg gagactacaa agatcatgac
     3061 attgattaca aggacgacga tgacaaataa gctagcttga ctgactgaga tacagcgtac
     3121 cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     3181 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     3241 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     3301 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca
     3361 tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     3421 tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc
     3481 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     3541 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta
     3601 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac
     3661 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct
     3721 tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg
     3781 ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc
     3841 cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg
     3901 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3961 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4021 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4081 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4141 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4201 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4261 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4321 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4381 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4441 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4501 agaaaaaaag gatctcaaga agat
//
