LOCUS       sequence-300312-        6272 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF7b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6272)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6272)
  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-m4252865
FEATURES             Location/Qualifiers
     source          1..6272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             134..154
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             155..181
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    234..355
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(245..264)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     299..318
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      470..516
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      546..682
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    727..798
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1024..1612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1104..1123)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2112..3321
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3158..3177
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3325..3924
                     /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(3325..3344)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3821..3841
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3951..3999
                     /note="synthetic polyadenylation signal"
     CDS             join(4265..4333,4334..5125)
                     /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(4483..4502)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5138..5274
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5426..5497
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5502..5548)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5566..5614
                     /note="synthetic polyadenylation signal"
     enhancer        5695..5998
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6000..6202
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6153..6173
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgata gaactatcgc tcatcgactt ttatctgtgc ttcttggctt tcctgctttt
       61 tctggtgctc attatgctaa tcattttttg gttcagtctg gagttacaag atcataatga
      121 gacatgtcac gccgaaaacc tttactttca gtcgtacccc tatgacgtac cagattatgc
      181 gtaagctagc ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata
      241 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
      301 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
      361 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
      421 caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc
      481 cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc
      541 taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
      601 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
      661 cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga
      721 tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca
      781 aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag
      841 tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt
      901 cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta
      961 aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1021 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1081 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1141 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1201 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1261 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1321 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1381 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1441 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1501 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1561 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1621 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1681 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1741 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1801 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc
     1861 gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg
     1921 ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt
     1981 ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga
     2041 gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc
     2101 ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac
     2161 ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc cgcccggacg
     2221 ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag
     2281 gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga
     2341 acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt
     2401 gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg
     2461 ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg
     2521 ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg
     2581 gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt
     2641 ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg
     2701 cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc
     2761 ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg ggcaccatct
     2821 ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg
     2881 ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag cgcgcgccct
     2941 cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg ccaccagccg
     3001 gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct agggtaggct
     3061 ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag gcgtcagttt
     3121 ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt ttgaactatg
     3181 cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga aatgtaatca
     3241 tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa attctggccg
     3301 tttttggctt ttttgttaga caacatgacc gagtacaagc ccacggtgcg cctcgccacc
     3361 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     3421 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     3481 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     3541 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     3601 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggagggcctc
     3661 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     3721 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggccgccgag
     3781 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     3841 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     3901 atgacccgca agcccggtgc ctaggctagt atgtaagcct agtcttagat aataaaatcg
     3961 ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg
     4021 tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg cttggcagtc
     4081 agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt gtactcaagc
     4141 ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg atcggtcaac
     4201 gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag caggaggctt
     4261 tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4321 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4381 tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4441 ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4501 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4561 cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4621 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4681 gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4741 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4801 gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct
     4861 agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct
     4921 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4981 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5041 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5101 tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc
     5161 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     5221 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatccc
     5281 gaatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac
     5341 agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct
     5401 ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc
     5461 cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc
     5521 cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa
     5581 cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca
     5641 gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     5701 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     5761 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     5821 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     5881 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     5941 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct
     6001 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     6061 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     6121 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     6181 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct
     6241 accatccact cgacacaccc gccagcggcc gc
//
