LOCUS       sequence-300694-        6314 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 6314)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6314)
  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-m4252834
FEATURES             Location/Qualifiers
     source          1..6314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(29..820,821..889))
                     /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     652..671
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1155..1203
                     /note="synthetic polyadenylation signal"
     CDS             complement(1230..1829)
                     /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(1313..1333)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1810..1829
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1833..3042)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(1977..1996)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      3542..4130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4031..4050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4356..4427
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4472..4608)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(4638..4684)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4799..4920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4836..4855)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4890..4909
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(4973..5038)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(5042..5062)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(5266..5468)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5295..5315)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5470..5773
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    5854..5902
                     /note="synthetic polyadenylation signal"
     terminator      5920..5966
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    5971..6042
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(join(6194..6314,1..16))
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 ttttctgcgc caatgcacct agaatcggtt accaatgctt aatcagtgag gcacctatct
       61 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
      121 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
      181 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
      241 gtcctgcaac tttatccgcc tccatccagt ctatcaactg ttgccgggaa gctagagtaa
      301 gtagttcgcc agttaatagt ttacgcaagg ttgttgccat tgctacaggc atcgtggtgt
      361 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
      421 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctcca atcgttgtca
      481 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
      541 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga atactcaacc aagtcattct
      601 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
      661 cgccacatag cagaacttta aaagtgctca tcattggaaa gcgttcttcg gggcgaaaac
      721 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgc gcacccaact
      781 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
      841 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcatg cgaaagcctc
      901 ctgctgatcc tgtctcttga tagatgtttg caaaagctag gactcgatta tacgttgacc
      961 gatcagacta tcaatatcta ggtttaaaca gatcggtacg ctgccgtgac caggcttgag
     1021 tacaatatct gacgattgga cttcctgggt cccaagctgg attccgactg gctgactgcc
     1081 aagcaacgat atggatcgtc gagactcgca ctgagaggat cgttctcggt acaccaggcg
     1141 cgcctagcgt tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt
     1201 attatctaag actaggctta catactagcc taggcaccgg gcttgcgggt catgcaccag
     1261 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
     1321 aaggggaggt tgcggggcgc ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcg
     1381 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag
     1441 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggccc
     1501 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
     1561 tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg
     1621 ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct
     1681 tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg
     1741 tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg
     1801 aggcgcaccg tgggcttgta ctcggtcatg ttgtctaaca aaaaagccaa aaacggccag
     1861 aatttagcgg acaatttaca agtctaacac tgaaaattac atattgaccc aaatgattac
     1921 atttcaaaag gtgcctaaaa aacttcacaa aacacactcg ccaaccccga gcgcatagtt
     1981 caaaaccgga gcttcagcta cttaagaaga taggtacata aaaccgacca aagaaactga
     2041 cgcctcactt atccctcccc tcaccagagg tccggcgcct gtcgattcag gagagcctac
     2101 cctaggcccg aaccctgcgt cctgcgacgg agaaaagcct accgcacacc taccggctgg
     2161 tggccccacc ctgcattata agccaacaga acgggtgacg tcacgacacg acgagggcgc
     2221 gcgctcccaa aggtacgggt gcactgccca acggcaccgc cataactgcc gcccccgcaa
     2281 cagacgacaa accgagttct ccagtcagtg acaaacttca cgtcagggtc cccagatggt
     2341 gcccagccca tctcacccga ataagagctt tcccgcatta gcgaagggct caagaccttg
     2401 ggttcttgcc gcccaccatg ccccccacct tgtttcaacg acctcacagc ccgcctcaca
     2461 agcgtcttcc attcaagact cgggaacagc cgccattttg ctgcgctccc cccaaccccc
     2521 agttcagggc aaccttgctc gcggacccag actacagccc ttggcggtct ctccacacgc
     2581 ttccgtccca ccgagcggcc cggcggccac gaaagccccg gccagcccag cagcccgcta
     2641 ctcaccaagt gacgatcaca gcgatccaca aacaagaacc gcgacccaaa tcccggctgc
     2701 gacggaacta gctgtgccac acccggcgcg tccttatata atcatcggcg ttcaccgccc
     2761 cacggagatc cctccgcaga atcgccgaga agggactact tttcctcgcc tgttccgctc
     2821 tctggaaaga aaaccagtgc cctagagtca cccaagtccc gtcctaaaat gtccttctgc
     2881 tgatactggg gttctaaggc cgagtcttat gagcagcggg ccgctgtcct gagcgtccgg
     2941 gcggaaggat caggacgctc ctgcgccctt cgtctgacgt ggcagcgctc gccgtgacga
     3001 ggggggcgcc cgcgggaggc gccaaaaccc ggcgcggagg ccaggcgcgc cccgatttcg
     3061 atattcctcg gatctcggat agatcgggat tgaggatgta ggttgaactt ctctcgggtg
     3121 atcccgatct cggatattcg ggattggagt aatacctggc gataggcgct gccaagcaat
     3181 agccaaggcg caaggccggc caagagactt gcgatagttc gttcgagagg tagcctgcaa
     3241 gatggatcgg acgaaaggtt tgaagatttt ccgacgaggg ccttggagag ttcggaaatt
     3301 taaatgcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3361 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3421 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3481 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3541 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3601 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3661 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3721 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3781 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3841 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3901 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3961 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4021 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4081 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4141 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg atttaaatga
     4201 aactcgtaat cggagaacgg gtgcaatagc aaggaccaga tcgaacttga ctgacgcaat
     4261 tctgagcgta gtcgaaaccc agacaagtcg atctcgccaa gtagctgact tgactggtta
     4321 attaagtttc gatagcccaa ggtaaccaat gagattaggg agggaagaaa gtttgatgaa
     4381 ctacttctga cccctagtgg tgtccagaaa agaccattaa aggaatgtct tgatcgaaat
     4441 cgagagatcg cctgcaggat agctgacgac cttaaggata aatttctggt aaggaggaca
     4501 cgtatggaag tgggcaagtt ggggaagccg tatccgttgc tgaatctggc atatgtggga
     4561 gtataagacg cgcagcgtcg catcaggcat ttttttctgc gccaatgccg gtagatagca
     4621 atccggcccg aggggcacaa aaaacccctc aagacccgtt tagaggcccc aaggggttat
     4681 gctacgatac cgatagagat gggccaatac cacatttgta gaggttttac ttgctttaaa
     4741 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     4801 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4861 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4921 tcatgtctgt gagctgaagg tacgctgtat ctcagtcagt caagctagct tatttgtcat
     4981 cgtcgtcctt gtaatcaatg tcatgatctt tgtagtctcc atcgtggtcc ttataatcca
     5041 tcgactgaaa gtaaaggttt tcggcgtgac aagtttcatt atgatcttgc agttcaagtg
     5101 agaaccaaaa gataataagc ataattaaaa caaggaatag cagaaaggct aaaaagcaca
     5161 aatagaagtc aattaatgaa agttcaatca tggtggcggc cgctggcggg tgtgtcgagt
     5221 ggatggtagg atcgacaaag atctgatctg acggttcact aaaccagctc tgcttatata
     5281 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     5341 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     5401 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     5461 ccgcatcagc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     5521 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     5581 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     5641 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     5701 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     5761 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     5821 ctagatctgg ctacactcga tcagcagtta gataataaaa tcgctatcca tcgaagatgg
     5881 atgtgtgttg gttttttgtg tgtgtaacgc aacgattgat agcataaccc cttggggcct
     5941 ctaaacgggt cttgaggggt tttttgaatt gagaggtagc tgaagctgct gttatgacca
     6001 ctagagggaa gaagatacct gtggagctaa tggtccaaga tgtcgggagc cgcgatcgct
     6061 gccaaaaagg taagctgacg atagctgacg agattcgatg cccgacgagc aaatgagcgc
     6121 cacggctgtt cgcgtaagct cgaaacgacg gagattcgat agagccagag tcgagtttaa
     6181 acgattcggg atcttaagga taaatttctg gtaaggagga cacgtatgga agtgggcaag
     6241 ttggggaagc cgtatccgtt gctgaatctg gcatatgtgg gagtataaga cgcgcagcgt
     6301 cgcatcaggc attt
//
