LOCUS       sequence-297319-        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             251..271
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             272..289
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    342..463
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(353..372)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     407..426
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      578..624
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      654..790
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    835..906
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1132..1720)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1212..1231)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2466..2534,2535..3326)
                     /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(2684..2703)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3339..3475
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3627..3698
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3703..3749)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3767..3815
                     /note="synthetic polyadenylation signal"
     enhancer        3896..4199
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4201..4403
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4354..4374
                     /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 gcaccatcac catcaccatt aagctagctt
      301 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt
      361 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
      421 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
      481 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
      541 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct
      601 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg
      661 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
      721 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
      781 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc
      841 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc
      901 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta
      961 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg
     1021 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc
     1081 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1141 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1201 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1261 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1321 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1381 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1441 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1501 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1561 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1621 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1681 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1741 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1801 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1861 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1921 ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag
     1981 gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa
     2041 cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat
     2101 cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac
     2161 ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg
     2221 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt
     2281 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag
     2341 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa
     2401 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct
     2461 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2521 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2581 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2641 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2701 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2761 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2821 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2881 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2941 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3001 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc
     3061 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc
     3121 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3181 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3241 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3301 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc
     3361 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     3421 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc
     3481 cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa
     3541 cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc
     3601 tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt
     3661 ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac
     3721 ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca
     3781 acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc
     3841 agatctagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     3901 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     3961 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     4021 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     4081 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     4141 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
     4201 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     4261 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     4321 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     4381 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc
     4441 taccatccac tcgacacacc cgccagcggc cgc
//
