LOCUS       sequence-297339-        4356 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 4356)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4356)
  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-m4133204
FEATURES             Location/Qualifiers
     source          1..4356
                     /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"
     CDS             join(2349..2417,2418..3209)
                     /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(2567..2586)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3222..3358
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3510..3581
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3586..3632)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3650..3698
                     /note="synthetic polyadenylation signal"
     enhancer        3779..4082
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4084..4286
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4237..4257
                     /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 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     2161 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     2221 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     2281 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     2341 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2401 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2461 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2521 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2581 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2641 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2701 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2761 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2821 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2881 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     2941 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     3001 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3061 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3121 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3181 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     3241 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     3301 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     3361 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc
     3421 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt
     3481 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt
     3541 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
     3601 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa
     3661 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta
     3721 gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     3781 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     3841 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     3901 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     3961 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     4021 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     4081 tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     4141 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     4201 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     4261 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga
     4321 tcctaccatc cactcgacac acccgccagc ggccgc
//
