LOCUS       sequence-297337-        6290 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 6290)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6290)
  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-m4252856
FEATURES             Location/Qualifiers
     source          1..6290
                     /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"
     promoter        2130..3339
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3176..3195
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3343..3942
                     /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(3343..3362)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3839..3859
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3969..4017
                     /note="synthetic polyadenylation signal"
     CDS             join(4283..4351,4352..5143)
                     /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(4501..4520)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5156..5292
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5444..5515
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5520..5566)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5584..5632
                     /note="synthetic polyadenylation signal"
     enhancer        5713..6016
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6018..6220
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6171..6191
                     /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 gcctccgcgc cgggttttgg cgcctcccgc
     2161 gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct
     2221 gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa
     2281 ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt
     2341 ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag
     2401 ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag
     2461 ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt
     2521 cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg
     2581 gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt
     2641 tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa
     2701 tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg
     2761 cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag
     2821 atgggctggg caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg
     2881 tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc
     2941 tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca
     3001 gggtggggcc accagccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt
     3061 tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga
     3121 taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag
     3181 ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca
     3241 ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg
     3301 tccgctaaat tctggccgtt tttggctttt ttgttagaca acatgaccga gtacaagccc
     3361 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     3421 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     3481 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     3541 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     3601 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     3661 caacagatgg agggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     3721 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     3781 ggagtggagg ccgccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     3841 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     3901 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct aggctagtat gtaagcctag
     3961 tcttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt
     4021 aacgctaggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca
     4081 tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt
     4141 cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg
     4201 atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac
     4261 aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4321 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4381 tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa
     4441 gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct
     4501 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4561 acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga
     4621 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4681 caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat
     4741 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4801 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac
     4861 tggcgaacta cttactctag cttcccggca acagttgata gactggatgg aggcggataa
     4921 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4981 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5041 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5101 acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt
     5161 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     5221 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     5281 atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg
     5341 tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca
     5401 gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc
     5461 tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc
     5521 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt
     5581 acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact
     5641 gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata
     5701 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     5761 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     5821 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     5881 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     5941 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     6001 catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     6061 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     6121 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     6181 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     6241 cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
