LOCUS       sequence-297304-        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-m4134465
FEATURES             Location/Qualifiers
     source          1..4407
                     /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             158..223
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    276..397
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(287..306)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     341..360
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      512..558
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      588..724
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    769..840
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1066..1654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1146..1165)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2400..2468,2469..3260)
                     /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(2618..2637)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3273..3409
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3561..3632
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3637..3683)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3701..3749
                     /note="synthetic polyadenylation signal"
     enhancer        3830..4133
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4135..4337
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4288..4308
                     /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 gtcgatggat tataaggacc acgatggaga
      181 ctacaaagat catgacattg attacaagga cgacgatgac aaataagcta gcttgactga
      241 ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca
      301 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
      361 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
      421 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
      481 atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg
      541 gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga
      601 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
      661 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
      721 ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat
      781 ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta
      841 atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc
      901 gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg
      961 tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg
     1021 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1081 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1141 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1201 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1261 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1321 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1381 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1441 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1501 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1561 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1621 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1681 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1741 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1801 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1861 gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc
     1921 gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact
     1981 atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag
     2041 gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat
     2101 cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggtgtac
     2161 cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca
     2221 gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt
     2281 cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata
     2341 atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca
     2401 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2461 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc
     2521 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2581 aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2641 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2701 ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2761 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg
     2821 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2881 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2941 ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt
     3001 cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3061 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3121 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3181 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3241 cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg
     3301 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     3361 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc
     3421 gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg
     3481 tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg
     3541 gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct tcttccctct
     3601 agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt
     3661 agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac
     3721 atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct
     3781 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     3841 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     3901 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     3961 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     4021 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     4081 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc
     4141 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     4201 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     4261 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     4321 tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat
     4381 ccactcgaca cacccgccag cggccgc
//
