LOCUS       sequence-295219-        4728 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 4728)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4728)
  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-m4133871
FEATURES             Location/Qualifiers
     source          1..4728
                     /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"
     CDS             join(2721..2789,2790..3581)
                     /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(2939..2958)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3594..3730
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3882..3953
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3958..4004)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4022..4070
                     /note="synthetic polyadenylation signal"
     enhancer        4151..4454
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4456..4658
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4609..4629
                     /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 gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     2521 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     2581 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     2641 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     2701 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     2761 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2821 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2881 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     2941 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3001 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     3061 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3121 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3181 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3241 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     3301 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     3361 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3421 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3481 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3541 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     3601 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     3661 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     3721 ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt
     3781 tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc
     3841 tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc
     3901 cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa
     3961 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac
     4021 acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc
     4081 tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata
     4141 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     4201 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     4261 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     4321 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     4381 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     4441 tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     4501 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     4561 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     4621 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca
     4681 gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgc
//
