LOCUS       sequence-297324-        4383 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 4383)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4383)
  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-m4133176
FEATURES             Location/Qualifiers
     source          1..4383
                     /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..199
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    252..373
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(263..282)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     317..336
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      488..534
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      564..700
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    745..816
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1042..1630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1122..1141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2376..2444,2445..3236)
                     /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(2594..2613)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3249..3385
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3537..3608
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3613..3659)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3677..3725
                     /note="synthetic polyadenylation signal"
     enhancer        3806..4109
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4111..4313
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4264..4284
                     /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 gtcgaaagaa accgcggcag ctaagttcga
      181 gaggcagcac atggacagtt aagctagctt gactgactga gatacagcgt accttcagct
      241 cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
      301 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
      361 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
      421 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc
      481 ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc
      541 tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc
      601 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
      661 acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag
      721 gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg
      781 gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat
      841 cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga
      901 ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc
      961 gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1021 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1081 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1141 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1201 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1261 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1321 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1381 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1441 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1501 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1561 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1621 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1681 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1741 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1801 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1861 agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg
     1921 tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt
     1981 gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc
     2041 gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat
     2101 ccgaggaata tcgaaatcgg ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag
     2161 tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag
     2221 gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta
     2281 aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca
     2341 tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg
     2401 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2461 tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc
     2521 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca
     2581 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     2641 tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa
     2701 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2761 ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt
     2821 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     2881 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc
     2941 gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga
     3001 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3061 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3121 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3181 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga
     3241 ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     3301 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     3361 ctccttacca gaaatttatc cttaagatcc cgaatcgttt aaactcgact ctggctctat
     3421 cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca
     3481 tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc
     3541 ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc
     3601 tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat
     3661 caatcgttgc gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt
     3721 ttattatcta actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag
     3781 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     3841 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     3901 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     3961 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     4021 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     4081 tctacgtatt agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc
     4141 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     4201 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     4261 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
     4321 tgaaccgtca gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc
     4381 cgc
//
