LOCUS       sequence-297585-        6347 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 6347)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6347)
  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-m4252978
FEATURES             Location/Qualifiers
     source          1..6347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    309..430
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(320..339)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     374..393
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      545..591
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      621..757
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    802..873
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1099..1687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1179..1198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2187..3396
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3233..3252
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3400..3999
                     /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(3400..3419)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3896..3916
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4026..4074
                     /note="synthetic polyadenylation signal"
     CDS             join(4340..4408,4409..5200)
                     /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(4558..4577)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5213..5349
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5501..5572
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5577..5623)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5641..5689
                     /note="synthetic polyadenylation signal"
     enhancer        5770..6073
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6075..6277
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6228..6248
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagc aaaatgtctg acgtcaagtg tacatccgta gttctgttat cagtgcttca
       61 acagctgcgg gtggagagtt cgtcaaagct ctgggcccag tgcgtgcagc tgcacaatga
      121 tatattgctc gcgaaagata ctacggaagc tttcgagaag atggtcagct tgctatctgt
      181 tctactgagc atgcaaggag cagtagacat caacaaactt tgcgaagaga tgttagacaa
      241 ccgagccacc ctgcaataag ctagcttgac tgactgagat acagcgtacc ttcagctcac
      301 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
      361 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
      421 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
      481 ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt
      541 atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg
      601 ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
      661 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
      721 tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg
      781 atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc
      841 agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga
      901 aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta
      961 cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat
     1021 tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1081 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1141 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1201 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1261 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1321 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1381 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1441 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1501 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1561 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1621 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1681 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1741 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1801 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1861 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1921 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc
     1981 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg
     2041 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag
     2101 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg
     2161 aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg
     2221 cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat
     2281 ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc
     2341 cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc
     2401 tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga
     2461 tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta
     2521 gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac
     2581 ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg
     2641 ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc
     2701 cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg
     2761 aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg
     2821 caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc gggtgagatg
     2881 ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt
     2941 gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt
     3001 gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg
     3061 tggggccacc agccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg
     3121 ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa
     3181 gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc
     3241 cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct
     3301 tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg taaattgtcc
     3361 gctaaattct ggccgttttt ggcttttttg ttagacaaca tgaccgagta caagcccacg
     3421 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     3481 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     3541 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     3601 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     3661 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     3721 cagatggagg gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     3781 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     3841 gtggaggccg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     3901 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     3961 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctagg ctagtatgta agcctagtct
     4021 tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac
     4081 gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     4141 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     4201 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     4261 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     4321 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     4381 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4441 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4501 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     4561 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4621 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     4681 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4741 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4801 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4861 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     4921 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     4981 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5041 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5101 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5161 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     5221 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     5281 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     5341 tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     5401 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     5461 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     5521 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     5581 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     5641 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     5701 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     5761 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     5821 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     5881 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     5941 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     6001 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     6061 cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     6121 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     6181 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     6241 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag
     6301 atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
