LOCUS       sequence-300197-        6263 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 6263)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6263)
  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-m4252870
FEATURES             Location/Qualifiers
     source          1..6263
                     /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..172
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    225..346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(236..255)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     290..309
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      461..507
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      537..673
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    718..789
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1015..1603)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1095..1114)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2103..3312
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3149..3168
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3316..3915
                     /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(3316..3335)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3812..3832
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3942..3990
                     /note="synthetic polyadenylation signal"
     CDS             join(4256..4324,4325..5116)
                     /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(4474..4493)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5129..5265
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5417..5488
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5493..5539)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5557..5605
                     /note="synthetic polyadenylation signal"
     enhancer        5686..5989
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5991..6193
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6144..6164
                     /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 gtcgcaccat caccatcacc attaagctag
      181 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
      241 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
      301 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
      361 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
      421 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
      481 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
      541 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
      601 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
      661 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
      721 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
      781 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
      841 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
      901 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
      961 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1021 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1081 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1141 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1201 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1261 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1321 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1381 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1441 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1501 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1561 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1621 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1681 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     1741 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     1801 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     1861 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     1921 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     1981 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     2041 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     2101 ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc
     2161 tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac gctcaggaca
     2221 gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta
     2281 ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag
     2341 gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga
     2401 tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg
     2461 tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc
     2521 tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg
     2581 ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc
     2641 gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg
     2701 aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgagccct
     2761 tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc tggggaccct
     2821 gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt
     2881 tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc
     2941 gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccaccagcc ggtaggtgtg
     3001 cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat
     3061 cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg
     3121 gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg
     3181 ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca
     3241 atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct
     3301 tttttgttag acaacatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     3361 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     3421 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     3481 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     3541 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     3601 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggagggcct cctggcgccg
     3661 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     3721 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggccgccga gcgcgccggg
     3781 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     3841 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     3901 aagcccggtg cctaggctag tatgtaagcc tagtcttaga taataaaatc gctatccatc
     3961 gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag
     4021 aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg
     4081 gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg
     4141 gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg
     4201 agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag
     4261 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4321 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt
     4381 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4441 acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4501 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4561 gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4621 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg
     4681 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4741 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4801 agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg
     4861 gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4921 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4981 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5041 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5101 gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac
     5161 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     5221 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt
     5281 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     5341 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     5401 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     5461 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     5521 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     5581 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     5641 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     5701 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     5761 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     5821 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     5881 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     5941 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
     6001 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     6061 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     6121 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     6181 tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac
     6241 tcgacacacc cgccagcggc cgc
//
