LOCUS       sequence-294998-        6628 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 6628)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6628)
  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-m4133246
FEATURES             Location/Qualifiers
     source          1..6628
                     /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             503..520
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    597..718
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(608..627)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     662..681
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      833..879
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      909..1045
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1090..1161
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1387..1975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1467..1486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2475..3684
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3521..3540
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3688..4287
                     /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(3688..3707)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4184..4204
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4314..4362
                     /note="synthetic polyadenylation signal"
     CDS             join(4628..4696,4697..5488)
                     /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(4846..4865)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5501..5637
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5782..5853
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5858..5904)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5922..5970
                     /note="synthetic polyadenylation signal"
     enhancer        6051..6354
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6356..6558
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6509..6529
                     /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 gtcgatgagt gacaaaatca ttcatctaac
      181 cgatgactcc tttgatacag acgtgttgaa ggccgatggt gcgatactgg tagatttctg
      241 ggctgaatgg tgtggcccgt gcaagatgat cgcacccatt ctggacgaga tcgccgacga
      301 gtatcaagga aaattaactg tcgctaagct taatatagat cagaacccag ggacggcacc
      361 taaatacggc atcaggggaa tacctaccct cctgctcttt aagaatgggg aagttgccgc
      421 gacaaaagtg ggagccctgt ctaagggcca gctaaaagag ttcttggacg ctaacctggc
      481 aggttcaggc tcggggcaca tgcaccatca ccatcaccat agcagcgggc tcgtgccacg
      541 cggctaagct agcttgactg actgagatac agcgtacctt cagctcacag acatgataag
      601 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
      661 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
      721 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
      781 aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa
      841 ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct
      901 atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
      961 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     1021 ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt
     1081 cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca
     1141 tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac
     1201 cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg
     1261 cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat
     1321 ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1381 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1441 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1501 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1561 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1621 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1681 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1741 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1801 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1861 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1921 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1981 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2041 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2101 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2161 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt
     2221 tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc
     2281 aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg
     2341 cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc
     2401 cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa
     2461 atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt
     2521 cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg
     2581 acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag
     2641 aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc
     2701 ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc
     2761 ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag
     2821 ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc
     2881 gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt
     2941 gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg
     3001 gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg
     3061 aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag
     3121 gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctgggcacca
     3181 tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc
     3241 gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc
     3301 cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccacctg
     3361 ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag
     3421 gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag
     3481 tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact
     3541 atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa
     3601 tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc taaattctgg
     3661 ccgtttttgg cttttttgtt agacaacatg accgagtaca agcccacggt gcgcctcgcc
     3721 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     3781 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     3841 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     3901 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     3961 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggagggc
     4021 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     4081 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggccgcc
     4141 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     4201 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     4261 tgcatgaccc gcaagcccgg tgcctaggct agtatgtaag cctagtctta gataataaaa
     4321 tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc
     4381 tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg ttgcttggca
     4441 gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat attgtactca
     4501 agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt ctgatcggtc
     4561 aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat cagcaggagg
     4621 ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4681 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4741 gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4801 tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4861 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4921 tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4981 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5041 aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5101 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5161 cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg aactacttac
     5221 tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg caggaccact
     5281 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5341 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5401 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5461 aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc agaaaaaaat
     5521 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     5581 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaagat
     5641 cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc
     5701 gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt
     5761 ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc
     5821 tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt
     5881 tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca
     5941 catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc
     6001 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     6061 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     6121 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     6181 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     6241 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     6301 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg
     6361 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     6421 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     6481 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     6541 gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca
     6601 tccactcgac acacccgcca gcggccgc
//
