LOCUS       sequence-299525-        4416 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 4416)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4416)
  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-m4133519
FEATURES             Location/Qualifiers
     source          1..4416
                     /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..232
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    285..406
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(296..315)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     350..369
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      521..567
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      597..733
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    778..849
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1075..1663)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1155..1174)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2409..2477,2478..3269)
                     /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(2627..2646)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3282..3418
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3570..3641
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3646..3692)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3710..3758
                     /note="synthetic polyadenylation signal"
     enhancer        3839..4142
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4144..4346
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4297..4317
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgatt gaactttcat taattgactt ctatttgtgc tttttagcct ttctgctatt
       61 ccttgtttta attatgctta ttatcttttg gttctcactt gaactgcaag atcataatga
      121 aacttgtcac gccgaaaacc tttactttca gtcgaagagg cgatggaaaa agaattttat
      181 cgcggtgagt gcagccaaca gattcaaaaa gatatcctcg tctggtgccc tctaagctag
      241 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
      301 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
      361 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
      421 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
      481 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
      541 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
      601 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
      661 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
      721 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
      781 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
      841 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
      901 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
      961 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
     1021 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1081 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1141 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1201 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1261 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1321 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1381 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1441 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1501 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1561 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1621 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1681 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1741 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     1801 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     1861 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     1921 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     1981 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     2041 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     2101 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     2161 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     2221 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     2281 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     2341 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     2401 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2461 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2521 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2581 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2641 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2701 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2761 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2821 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2881 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2941 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     3001 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     3061 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3121 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3181 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3241 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     3301 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     3361 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     3421 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc
     3481 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt
     3541 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt
     3601 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
     3661 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa
     3721 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta
     3781 gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     3841 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     3901 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     3961 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     4021 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     4081 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     4141 tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     4201 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     4261 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     4321 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga
     4381 tcctaccatc cactcgacac acccgccagc ggccgc
//
