LOCUS       sequence-300603-        6759 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 6759)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6759)
  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-m4252967
FEATURES             Location/Qualifiers
     source          1..6759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    31..79
                     /note="synthetic polyadenylation signal"
     terminator      97..143
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    148..219
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(371..507)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(520..1311,1312..1380))
                     /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     1143..1162
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1646..1694
                     /note="synthetic polyadenylation signal"
     CDS             complement(1721..2749)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2753..3962)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2897..2916)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4462..5050
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4951..4970
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5276..5347
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5392..5528)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5558..5604)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5719..5840
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5756..5775)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5810..5829
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5909..5974)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(5978..5998)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(6202..6404)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6231..6251)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6406..6709
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gatctggcta cactcgatca gcagttagat aataaaatcg ctatccatcg aagatggatg
       61 tgtgttggtt ttttgtgtgt gtaacgcaac gattgatagc ataacccctt ggggcctcta
      121 aacgggtctt gaggggtttt ttgaattgag aggtagctga agctgctgtt atgaccacta
      181 gagggaagaa gatacctgtg gagctaatgg tccaagatgt cgggagccgc gatcgctgcc
      241 aaaaaggtaa gctgacgata gctgacgaga ttcgatgccc gacgagcaaa tgagcgccac
      301 ggctgttcgc gtaagctcga aacgacggag attcgataga gccagagtcg agtttaaacg
      361 attcgggatc ttaaggataa atttctggta aggaggacac gtatggaagt gggcaagttg
      421 gggaagccgt atccgttgct gaatctggca tatgtgggag tataagacgc gcagcgtcgc
      481 atcaggcatt tttttctgcg ccaatgcacc tagaatcggt taccaatgct taatcagtga
      541 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      601 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      661 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      721 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctatcaact gttgccggga
      781 agctagagta agtagttcgc cagttaatag tttacgcaag gttgttgcca ttgctacagg
      841 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
      901 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
      961 aatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1021 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg aatactcaac
     1081 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1141 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa agcgttcttc
     1201 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1261 cgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1321 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1381 gcgaaagcct cctgctgatc ctgtctcttg atagatgttt gcaaaagcta ggactcgatt
     1441 atacgttgac cgatcagact atcaatatct aggtttaaac agatcggtac gctgccgtga
     1501 ccaggcttga gtacaatatc tgacgattgg acttcctggg tcccaagctg gattccgact
     1561 ggctgactgc caagcaacga tatggatcgt cgagactcgc actgagagga tcgttctcgg
     1621 tacaccaggc gcgcctagcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg
     1681 atagcgattt tattatctaa gactaggctt acatactagc ttattccttt gccctcggac
     1741 gagtgctggg gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga
     1801 cggcggcgct tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga
     1861 ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct
     1921 gatagagttg gtcaagacca atgcggagca tatacgcccg gagccggggc gatcctgcaa
     1981 gctccggatg cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc
     2041 tccagaagaa gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt
     2101 caatgaccgc tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt
     2161 gcacgaggtg ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct
     2221 gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat
     2281 cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga
     2341 atgggccgaa cccgctcgtc tggctaagat cggcggcagc gatggcatcc attgcctccg
     2401 cgaccggctg cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct
     2461 gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa ctccccaatg tcaagcactt
     2521 ccggaatcgg gagcgcggcg gatgcaaagt gccgataaac ataacgatct ttgtagaaac
     2581 catcggcgca gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag
     2641 cacgagattc ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga
     2701 tcagaaactt ctcgacagac gtcgcggtga gttcaggctt tttacccatg ttgtctaaca
     2761 aaaaagccaa aaacggccag aatttagcgg acaatttaca agtctaacac tgaaaattac
     2821 atattgaccc aaatgattac atttcaaaag gtgcctaaaa aacttcacaa aacacactcg
     2881 ccaaccccga gcgcatagtt caaaaccgga gcttcagcta cttaagaaga taggtacata
     2941 aaaccgacca aagaaactga cgcctcactt atccctcccc tcaccagagg tccggcgcct
     3001 gtcgattcag gagagcctac cctaggcccg aaccctgcgt cctgcgacgg agaaaagcct
     3061 accgcacacc taccggctgg tggccccacc ctgcattata agccaacaga acgggtgacg
     3121 tcacgacacg acgagggcgc gcgctcccaa aggtacgggt gcactgccca acggcaccgc
     3181 cataactgcc gcccccgcaa cagacgacaa accgagttct ccagtcagtg acaaacttca
     3241 cgtcagggtc cccagatggt gcccagccca tctcacccga ataagagctt tcccgcatta
     3301 gcgaagggct caagaccttg ggttcttgcc gcccaccatg ccccccacct tgtttcaacg
     3361 acctcacagc ccgcctcaca agcgtcttcc attcaagact cgggaacagc cgccattttg
     3421 ctgcgctccc cccaaccccc agttcagggc aaccttgctc gcggacccag actacagccc
     3481 ttggcggtct ctccacacgc ttccgtccca ccgagcggcc cggcggccac gaaagccccg
     3541 gccagcccag cagcccgcta ctcaccaagt gacgatcaca gcgatccaca aacaagaacc
     3601 gcgacccaaa tcccggctgc gacggaacta gctgtgccac acccggcgcg tccttatata
     3661 atcatcggcg ttcaccgccc cacggagatc cctccgcaga atcgccgaga agggactact
     3721 tttcctcgcc tgttccgctc tctggaaaga aaaccagtgc cctagagtca cccaagtccc
     3781 gtcctaaaat gtccttctgc tgatactggg gttctaaggc cgagtcttat gagcagcggg
     3841 ccgctgtcct gagcgtccgg gcggaaggat caggacgctc ctgcgccctt cgtctgacgt
     3901 ggcagcgctc gccgtgacga ggggggcgcc cgcgggaggc gccaaaaccc ggcgcggagg
     3961 ccaggcgcgc cccgatttcg atattcctcg gatctcggat agatcgggat tgaggatgta
     4021 ggttgaactt ctctcgggtg atcccgatct cggatattcg ggattggagt aatacctggc
     4081 gataggcgct gccaagcaat agccaaggcg caaggccggc caagagactt gcgatagttc
     4141 gttcgagagg tagcctgcaa gatggatcgg acgaaaggtt tgaagatttt ccgacgaggg
     4201 ccttggagag ttcggaaatt taaatgcaac tatggatgaa cgaaatagac agatcgctga
     4261 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4321 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4381 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4441 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4501 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4561 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4621 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4681 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4741 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4801 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4861 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4921 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4981 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5041 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5101 tgctcacatg atttaaatga aactcgtaat cggagaacgg gtgcaatagc aaggaccaga
     5161 tcgaacttga ctgacgcaat tctgagcgta gtcgaaaccc agacaagtcg atctcgccaa
     5221 gtagctgact tgactggtta attaagtttc gatagcccaa ggtaaccaat gagattaggg
     5281 agggaagaaa gtttgatgaa ctacttctga cccctagtgg tgtccagaaa agaccattaa
     5341 aggaatgtct tgatcgaaat cgagagatcg cctgcaggat agctgacgac cttaaggata
     5401 aatttctggt aaggaggaca cgtatggaag tgggcaagtt ggggaagccg tatccgttgc
     5461 tgaatctggc atatgtggga gtataagacg cgcagcgtcg catcaggcat ttttttctgc
     5521 gccaatgccg gtagatagca atccggcccg aggggcacaa aaaacccctc aagacccgtt
     5581 tagaggcccc aaggggttat gctacgatac cgatagagat gggccaatac cacatttgta
     5641 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     5701 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5761 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5821 aaactcatca atgtatctta tcatgtctgt gagctgaagg tacgctgtat ctcagtcagt
     5881 caagctagca gaggcctgcg gatccttatt tgtcatcgtc gtccttgtaa tcaatgtcat
     5941 gatctttgta gtctccatcg tggtccttat aatccatcga ctgaaagtaa aggttttcgg
     6001 cgtgacatgt ctcattatga tcttgtaact ccagactgaa ccaaaaaatg attagcataa
     6061 tgagcaccag aaaaagcagg aaagccaaga agcacagata aaagtcgatg agcgatagtt
     6121 ctatcatggt ggcggccgct ggcgggtgtg tcgagtggat ggtaggatcg acaaagatct
     6181 gatctgacgg ttcactaaac cagctctgct tatatagacc tcccaccgta cacgcctacc
     6241 gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg
     6301 gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc
     6361 aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcagcatgg taatagcgat
     6421 gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata
     6481 atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata
     6541 cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg
     6601 gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg
     6661 gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat
     6721 atatgggcta tgaactaatg accccgtaat tgattacta
//
