LOCUS       sequence-297598-        6224 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 6224)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6224)
  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-m4252973
FEATURES             Location/Qualifiers
     source          1..6224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    186..307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(197..216)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     251..270
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      422..468
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      498..634
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    679..750
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(976..1564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1056..1075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2064..3273
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3110..3129
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3277..3876
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3277..3296)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3773..3793
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3903..3951
                     /note="synthetic polyadenylation signal"
     CDS             join(4217..4285,4286..5077)
                     /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(4435..4454)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5090..5226
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5378..5449
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5454..5500)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5518..5566
                     /note="synthetic polyadenylation signal"
     enhancer        5647..5950
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5952..6154
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6105..6125
                     /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 gcctaagcta gcttgactga ctgagataca gcgtaccttc agctcacaga
      181 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
      241 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
      301 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
      361 ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc
      421 gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc
      481 cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
      541 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
      601 atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc
      661 tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga
      721 agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac
      781 ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc
      841 tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac
      901 gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      961 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1021 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1081 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1141 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1201 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1261 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1321 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1381 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1441 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1501 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1561 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1621 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1681 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1741 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1801 atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat
     1861 ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct
     1921 tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc
     1981 gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg
     2041 aatatcgaaa tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc
     2101 ccccctcgtc acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct
     2161 tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag
     2221 tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt
     2281 ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct
     2341 ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt
     2401 ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg
     2461 gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga
     2521 cggaagcgtg tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct
     2581 gaactggggg ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag
     2641 acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa
     2701 gaacccaagg tcttgagccc ttcgctaatg cgggaaagct cttattcggg tgagatgggc
     2761 tgggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc
     2821 gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg
     2881 agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg
     2941 ggccaccagc cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc
     3001 ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg
     3061 aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg
     3121 ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt
     3181 gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agacttgtaa attgtccgct
     3241 aaattctggc cgtttttggc ttttttgtta gacaacatga ccgagtacaa gcccacggtg
     3301 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     3361 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     3421 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     3481 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     3541 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     3601 atggagggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     3661 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     3721 gaggccgccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     3781 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     3841 cgcacctggt gcatgacccg caagcccggt gcctaggcta gtatgtaagc ctagtcttag
     3901 ataataaaat cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct
     3961 aggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt
     4021 tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata
     4081 ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc
     4141 tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc
     4201 agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4261 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4321 agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4381 tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg
     4441 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4501 ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat
     4561 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4621 acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4681 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4741 gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga
     4801 actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc
     4861 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4921 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4981 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5041 cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca
     5101 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     5161 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     5221 ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga
     5281 gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc
     5341 gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca
     5401 ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa
     5461 cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac
     5521 acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat
     5581 cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga
     5641 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     5701 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     5761 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     5821 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     5881 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     5941 tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     6001 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     6061 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     6121 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc
     6181 tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
