LOCUS       sequence-300691-        4893 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 4893)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4893)
  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-m4133309
FEATURES             Location/Qualifiers
     source          1..4893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      60..106
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    111..182
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(334..470)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(483..1274,1275..1343))
                     /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     1106..1125
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2089..2677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2578..2597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2903..2974
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3019..3155)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(3185..3231)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3346..3467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3383..3402)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3437..3456
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3916..3936)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(3964..4041)
                     /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"
     CDS             complement(4075..4095)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(4299..4501)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4328..4348)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4503..4806
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    join(4887..4893,1..42)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc aacgattgat
       61 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgaatt gagaggtagc
      121 tgaagctgct gttatgacca ctagagggaa gaagatacct gtggagctaa tggtccaaga
      181 tgtcgggagc cgcgatcgct gccaaaaagg taagctgacg atagctgacg agattcgatg
      241 cccgacgagc aaatgagcgc cacggctgtt cgcgtaagct cgaaacgacg gagattcgat
      301 agagccagag tcgagtttaa acgattcggg atcttaagga taaatttctg gtaaggagga
      361 cacgtatgga agtgggcaag ttggggaagc cgtatccgtt gctgaatctg gcatatgtgg
      421 gagtataaga cgcgcagcgt cgcatcaggc atttttttct gcgccaatgc acctagaatc
      481 ggttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
      541 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
      601 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
      661 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
      721 cagtctatca actgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttacgc
      781 aaggttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
      841 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
      901 gcggttagct ccttcggtcc tccaatcgtt gtcagaagta agttggccgc agtgttatca
      961 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1021 tctgtgactg gtgaatactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     1081 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     1141 ctcatcattg gaaagcgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     1201 tccagttcga tgtaacccac tcgcgcaccc aactgatctt cagcatcttt tactttcacc
     1261 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     1321 acacggaaat gttgaatact catgcgaaag cctcctgctg atcctgtctc ttgatagatg
     1381 tttgcaaaag ctaggactcg attatacgtt gaccgatcag actatcaata tctaggttta
     1441 aacagatcgg tacgctgccg tgaccaggct tgagtacaat atctgacgat tggacttcct
     1501 gggtcccaag ctggattccg actggctgac tgccaagcaa cgatatggat cgtcgagact
     1561 cgcactgaga ggatcgttct cggtacacca ggcgcgcccc gatttcgata ttcctcggat
     1621 ctcggataga tcgggattga ggatgtaggt tgaacttctc tcgggtgatc ccgatctcgg
     1681 atattcggga ttggagtaat acctggcgat aggcgctgcc aagcaatagc caaggcgcaa
     1741 ggccggccaa gagacttgcg atagttcgtt cgagaggtag cctgcaagat ggatcggacg
     1801 aaaggtttga agattttccg acgagggcct tggagagttc ggaaatttaa atgcaactat
     1861 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1921 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1981 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2041 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2101 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2161 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2221 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2281 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2341 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2401 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2461 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2521 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2581 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2641 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2701 acggttcctg gccttttgct ggccttttgc tcacatgatt taaatgaaac tcgtaatcgg
     2761 agaacgggtg caatagcaag gaccagatcg aacttgactg acgcaattct gagcgtagtc
     2821 gaaacccaga caagtcgatc tcgccaagta gctgacttga ctggttaatt aagtttcgat
     2881 agcccaaggt aaccaatgag attagggagg gaagaaagtt tgatgaacta cttctgaccc
     2941 ctagtggtgt ccagaaaaga ccattaaagg aatgtcttga tcgaaatcga gagatcgcct
     3001 gcaggatagc tgacgacctt aaggataaat ttctggtaag gaggacacgt atggaagtgg
     3061 gcaagttggg gaagccgtat ccgttgctga atctggcata tgtgggagta taagacgcgc
     3121 agcgtcgcat caggcatttt tttctgcgcc aatgccggta gatagcaatc cggcccgagg
     3181 ggcacaaaaa acccctcaag acccgtttag aggccccaag gggttatgct acgataccga
     3241 tagagatggg ccaataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3301 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3361 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3421 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtgag
     3481 ctgaaggtac gctgtatctc agtcagtcaa gctagcttat tgatggttcg cgtctacttt
     3541 cggggcctgg gcatcattga gctttttggc ttccgcaagc aggttggcac tctgtgatgg
     3601 atcgtctttt agggactgga taaatgcatt tctttgttcc tcattcagat taggaaggtg
     3661 caatatctcg tagaaagcat tctgctgctc cttgttaaac ttgttatcaa cttttggagc
     3721 ctgtgcgtca ttcagcttct tggcttcggc caggagattt gcagactgcg agggatcgtc
     3781 tttcaggctc tgaatgaaag cgttgcgttg ctcttcgttc agatttggta agtgaaggat
     3841 ctcatagaaa gcgttctgct gttctttatt aaacttgtta tcgacggcct cgtcatgttg
     3901 tgccaagcca aattcaccct ggaaatagag attctcggag gcggtcgtag gaatgtcgta
     3961 atctagggcg ccgcttgaac tgatcttttt gaaccggtta gccgcagaca ctgcaataaa
     4021 gttcttcttc catctccttt tccagggatt tatcagcccc gggattcgcc gtcccgactg
     4081 aaagtaaagg ttttcggcgt gacatgtctc attatgatct tgtaactcca gactgaacca
     4141 aaaaatgatt agcataatga gcaccagaaa aagcaggaaa gccaagaagc acagataaaa
     4201 gtcgatgagc gatagttcta tcatggtggc ggccgctggc gggtgtgtcg agtggatggt
     4261 aggatcgaca aagatctgat ctgacggttc actaaaccag ctctgcttat atagacctcc
     4321 caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga
     4381 aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt
     4441 ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc
     4501 agcatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag
     4561 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     4621 gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
     4681 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     4741 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     4801 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactagatc
     4861 tggctacact cgatcagcag ttagataata aaa
//
