LOCUS       sequence-307374-        4407 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 4407)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4407)
  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-m4134115
FEATURES             Location/Qualifiers
     source          1..4407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(105..307)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(134..154)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        309..612
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    693..741
                     /note="synthetic polyadenylation signal"
     terminator      759..805
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    810..881
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1033..1169)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(1182..1973,1974..2042))
                     /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     1805..1824
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2788..3376
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3277..3296
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3602..3673
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3718..3854)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(3884..3930)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4045..4166
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4082..4101)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4136..4155
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(4219..4284)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(4288..4308)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
ORIGIN
        1 atagaagtca attaatgaaa gttcaatcat ggtggcggcc gctggcgggt gtgtcgagtg
       61 gatggtagga tcgacaaaga tctgatctga cggttcacta aaccagctct gcttatatag
      121 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
      181 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
      241 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
      301 cgcatcagca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
      361 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
      421 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
      481 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
      541 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
      601 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
      661 tagatctggc tacactcgat cagcagttag ataataaaat cgctatccat cgaagatgga
      721 tgtgtgttgg ttttttgtgt gtgtaacgca acgattgata gcataacccc ttggggcctc
      781 taaacgggtc ttgaggggtt ttttgaattg agaggtagct gaagctgctg ttatgaccac
      841 tagagggaag aagatacctg tggagctaat ggtccaagat gtcgggagcc gcgatcgctg
      901 ccaaaaaggt aagctgacga tagctgacga gattcgatgc ccgacgagca aatgagcgcc
      961 acggctgttc gcgtaagctc gaaacgacgg agattcgata gagccagagt cgagtttaaa
     1021 cgattcggga tcttaaggat aaatttctgg taaggaggac acgtatggaa gtgggcaagt
     1081 tggggaagcc gtatccgttg ctgaatctgg catatgtggg agtataagac gcgcagcgtc
     1141 gcatcaggca tttttttctg cgccaatgca cctagaatcg gttaccaatg cttaatcagt
     1201 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1261 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1321 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1381 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctatcaa ctgttgccgg
     1441 gaagctagag taagtagttc gccagttaat agtttacgca aggttgttgc cattgctaca
     1501 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1561 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1621 ccaatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1681 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgaatactca
     1741 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1801 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaagcgttct
     1861 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1921 cgcgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1981 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     2041 atgcgaaagc ctcctgctga tcctgtctct tgatagatgt ttgcaaaagc taggactcga
     2101 ttatacgttg accgatcaga ctatcaatat ctaggtttaa acagatcggt acgctgccgt
     2161 gaccaggctt gagtacaata tctgacgatt ggacttcctg ggtcccaagc tggattccga
     2221 ctggctgact gccaagcaac gatatggatc gtcgagactc gcactgagag gatcgttctc
     2281 ggtacaccag gcgcgccccg atttcgatat tcctcggatc tcggatagat cgggattgag
     2341 gatgtaggtt gaacttctct cgggtgatcc cgatctcgga tattcgggat tggagtaata
     2401 cctggcgata ggcgctgcca agcaatagcc aaggcgcaag gccggccaag agacttgcga
     2461 tagttcgttc gagaggtagc ctgcaagatg gatcggacga aaggtttgaa gattttccga
     2521 cgagggcctt ggagagttcg gaaatttaaa tgcaactatg gatgaacgaa atagacagat
     2581 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2641 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2701 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2761 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2821 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2881 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     2941 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3001 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3061 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3121 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3181 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3241 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3301 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3361 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3421 gccttttgct cacatgattt aaatgaaact cgtaatcgga gaacgggtgc aatagcaagg
     3481 accagatcga acttgactga cgcaattctg agcgtagtcg aaacccagac aagtcgatct
     3541 cgccaagtag ctgacttgac tggttaatta agtttcgata gcccaaggta accaatgaga
     3601 ttagggaggg aagaaagttt gatgaactac ttctgacccc tagtggtgtc cagaaaagac
     3661 cattaaagga atgtcttgat cgaaatcgag agatcgcctg caggatagct gacgacctta
     3721 aggataaatt tctggtaagg aggacacgta tggaagtggg caagttgggg aagccgtatc
     3781 cgttgctgaa tctggcatat gtgggagtat aagacgcgca gcgtcgcatc aggcattttt
     3841 ttctgcgcca atgccggtag atagcaatcc ggcccgaggg gcacaaaaaa cccctcaaga
     3901 cccgtttaga ggccccaagg ggttatgcta cgataccgat agagatgggc caataccaca
     3961 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     4021 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     4081 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4141 ttgtccaaac tcatcaatgt atcttatcat gtctgtgagc tgaaggtacg ctgtatctca
     4201 gtcagtcaag ctagcttatt tgtcatcgtc gtccttgtaa tcaatgtcat gatctttgta
     4261 gtctccatcg tggtccttat aatccatcga ctgaaagtaa aggttttcgg cgtgacaagt
     4321 ttcattatga tcttgcagtt caagtgagaa ccaaaagata ataagcataa ttaaaacaag
     4381 gaatagcaga aaggctaaaa agcacaa
//
