LOCUS       sequence-297621-        6341 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 6341)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6341)
  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-m4252970
FEATURES             Location/Qualifiers
     source          1..6341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    303..424
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(314..333)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     368..387
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      539..585
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      615..751
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    796..867
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1093..1681)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1173..1192)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2181..3390
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3227..3246
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3394..3993
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3394..3413)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3890..3910
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4020..4068
                     /note="synthetic polyadenylation signal"
     CDS             join(4334..4402,4403..5194)
                     /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(4552..4571)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5207..5343
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5495..5566
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5571..5617)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5635..5683
                     /note="synthetic polyadenylation signal"
     enhancer        5764..6067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6069..6271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6222..6242
                     /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 taagctagct tgactgactg agatacagcg taccttcagc tcacagacat
      301 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
      361 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
      421 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
      481 tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta
      541 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg
      601 attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
      661 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
      721 cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct
      781 cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt
      841 agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta
      901 attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca
      961 gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag
     1021 tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1081 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1141 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1201 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1261 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1321 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1381 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1441 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1501 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1561 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1621 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1681 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1741 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     1801 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     1861 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt
     1921 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca
     1981 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg
     2041 ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg
     2101 atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat
     2161 atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc
     2221 cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc aggagcgtcc tgatccttcc
     2281 gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga accccagtat
     2341 cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt tttctttcca
     2401 gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga gggatctccg
     2461 tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca gctagttccg
     2521 tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg tcacttggtg
     2581 agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc ggtgggacgg
     2641 aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa
     2701 ctgggggttg gggggagcgc agcaaaatgg cggctgttcc cgagtcttga atggaagacg
     2761 cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa
     2821 cccaaggtct tgagcccttc gctaatgcgg gaaagctctt attcgggtga gatgggctgg
     2881 gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc
     2941 tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac ctttgggagc
     3001 gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc agggtggggc
     3061 caccagccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta
     3121 gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg ataagtgagg
     3181 cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa gctccggttt
     3241 tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa
     3301 atgtaatcat ttgggtcaat atgtaatttt cagtgttaga cttgtaaatt gtccgctaaa
     3361 ttctggccgt ttttggcttt tttgttagac aacatgaccg agtacaagcc cacggtgcgc
     3421 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     3481 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     3541 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     3601 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     3661 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     3721 gagggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     3781 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     3841 gccgccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     3901 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     3961 acctggtgca tgacccgcaa gcccggtgcc taggctagta tgtaagccta gtcttagata
     4021 ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg taacgctagg
     4081 cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc
     4141 ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg tcagatattg
     4201 tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt gatagtctga
     4261 tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga caggatcagc
     4321 aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4381 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4441 tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4501 gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc tgctatgtgg
     4561 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4621 tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg atggcatgac
     4681 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4741 tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4801 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4861 tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa ctggcgaact
     4921 acttactcta gcttcccggc aacagttgat agactggatg gaggcggata aagttgcagg
     4981 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5041 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5101 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5161 tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa
     5221 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
     5281 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
     5341 taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc gtttcgagct
     5401 tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc
     5461 agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt
     5521 atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc
     5581 ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca
     5641 aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga
     5701 gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat atatggagtt
     5761 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     5821 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     5881 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     5941 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     6001 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     6061 taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     6121 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     6181 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     6241 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tcagatcttt
     6301 gtcgatccta ccatccactc gacacacccg ccagcggccg c
//
