LOCUS       sequence-300601-        6735 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 6735)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6735)
  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-m4252953
FEATURES             Location/Qualifiers
     source          1..6735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1438..2026
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1927..1946
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2252..2323
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2368..2504)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(2534..2580)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    2695..2816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2732..2751)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2786..2805
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(2885..2929)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(2930..2950)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(3154..3356)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3183..3203)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3358..3661
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    3742..3790
                     /note="synthetic polyadenylation signal"
     terminator      3808..3854
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    3859..3930
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4082..4218)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(4231..5022,5023..5091))
                     /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     4854..4873
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5357..5405
                     /note="synthetic polyadenylation signal"
     CDS             complement(5432..6460)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(join(6464..6735,1..938))
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(6608..6627)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 ggcccgaacc ctgcgtcctg cgacggagaa aagcctaccg cacacctacc ggctggtggc
       61 cccaccctgc attataagcc aacagaacgg gtgacgtcac gacacgacga gggcgcgcgc
      121 tcccaaaggt acgggtgcac tgcccaacgg caccgccata actgccgccc ccgcaacaga
      181 cgacaaaccg agttctccag tcagtgacaa acttcacgtc agggtcccca gatggtgccc
      241 agcccatctc acccgaataa gagctttccc gcattagcga agggctcaag accttgggtt
      301 cttgccgccc accatgcccc ccaccttgtt tcaacgacct cacagcccgc ctcacaagcg
      361 tcttccattc aagactcggg aacagccgcc attttgctgc gctcccccca acccccagtt
      421 cagggcaacc ttgctcgcgg acccagacta cagcccttgg cggtctctcc acacgcttcc
      481 gtcccaccga gcggcccggc ggccacgaaa gccccggcca gcccagcagc ccgctactca
      541 ccaagtgacg atcacagcga tccacaaaca agaaccgcga cccaaatccc ggctgcgacg
      601 gaactagctg tgccacaccc ggcgcgtcct tatataatca tcggcgttca ccgccccacg
      661 gagatccctc cgcagaatcg ccgagaaggg actacttttc ctcgcctgtt ccgctctctg
      721 gaaagaaaac cagtgcccta gagtcaccca agtcccgtcc taaaatgtcc ttctgctgat
      781 actggggttc taaggccgag tcttatgagc agcgggccgc tgtcctgagc gtccgggcgg
      841 aaggatcagg acgctcctgc gcccttcgtc tgacgtggca gcgctcgccg tgacgagggg
      901 ggcgcccgcg ggaggcgcca aaacccggcg cggaggccag gcgcgccccg atttcgatat
      961 tcctcggatc tcggatagat cgggattgag gatgtaggtt gaacttctct cgggtgatcc
     1021 cgatctcgga tattcgggat tggagtaata cctggcgata ggcgctgcca agcaatagcc
     1081 aaggcgcaag gccggccaag agacttgcga tagttcgttc gagaggtagc ctgcaagatg
     1141 gatcggacga aaggtttgaa gattttccga cgagggcctt ggagagttcg gaaatttaaa
     1201 tgcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1261 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1321 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1381 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1441 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1501 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1561 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1621 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1681 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1741 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1801 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     1861 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     1921 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     1981 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2041 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgattt aaatgaaact
     2101 cgtaatcgga gaacgggtgc aatagcaagg accagatcga acttgactga cgcaattctg
     2161 agcgtagtcg aaacccagac aagtcgatct cgccaagtag ctgacttgac tggttaatta
     2221 agtttcgata gcccaaggta accaatgaga ttagggaggg aagaaagttt gatgaactac
     2281 ttctgacccc tagtggtgtc cagaaaagac cattaaagga atgtcttgat cgaaatcgag
     2341 agatcgcctg caggatagct gacgacctta aggataaatt tctggtaagg aggacacgta
     2401 tggaagtggg caagttgggg aagccgtatc cgttgctgaa tctggcatat gtgggagtat
     2461 aagacgcgca gcgtcgcatc aggcattttt ttctgcgcca atgccggtag atagcaatcc
     2521 ggcccgaggg gcacaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta
     2581 cgataccgat agagatgggc caataccaca tttgtagagg ttttacttgc tttaaaaaac
     2641 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2701 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2761 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     2821 gtctgtgagc tgaaggtacg ctgtatctca gtcagtcaag ctagcagagg cctgcggatc
     2881 cttaactgtc catgtgctgc ctctcgaact tagctgccgc ggtttctttc gactgaaagt
     2941 aaaggttttc ggcgtgacat gtctcattat gatcttgtaa ctccagactg aaccaaaaaa
     3001 tgattagcat aatgagcacc agaaaaagca ggaaagccaa gaagcacaga taaaagtcga
     3061 tgagcgatag ttctatcatg gtggcggccg ctggcgggtg tgtcgagtgg atggtaggat
     3121 cgacaaagat ctgatctgac ggttcactaa accagctctg cttatataga cctcccaccg
     3181 tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc
     3241 ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa
     3301 tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcagcat
     3361 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat
     3421 gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact
     3481 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca
     3541 ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt
     3601 caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac
     3661 gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact agatctggct
     3721 acactcgatc agcagttaga taataaaatc gctatccatc gaagatggat gtgtgttggt
     3781 tttttgtgtg tgtaacgcaa cgattgatag cataacccct tggggcctct aaacgggtct
     3841 tgaggggttt tttgaattga gaggtagctg aagctgctgt tatgaccact agagggaaga
     3901 agatacctgt ggagctaatg gtccaagatg tcgggagccg cgatcgctgc caaaaaggta
     3961 agctgacgat agctgacgag attcgatgcc cgacgagcaa atgagcgcca cggctgttcg
     4021 cgtaagctcg aaacgacgga gattcgatag agccagagtc gagtttaaac gattcgggat
     4081 cttaaggata aatttctggt aaggaggaca cgtatggaag tgggcaagtt ggggaagccg
     4141 tatccgttgc tgaatctggc atatgtggga gtataagacg cgcagcgtcg catcaggcat
     4201 ttttttctgc gccaatgcac ctagaatcgg ttaccaatgc ttaatcagtg aggcacctat
     4261 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4321 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4381 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4441 tggtcctgca actttatccg cctccatcca gtctatcaac tgttgccggg aagctagagt
     4501 aagtagttcg ccagttaata gtttacgcaa ggttgttgcc attgctacag gcatcgtggt
     4561 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4621 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc caatcgttgt
     4681 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4741 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gaatactcaa ccaagtcatt
     4801 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4861 cgcgccacat agcagaactt taaaagtgct catcattgga aagcgttctt cggggcgaaa
     4921 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gcgcacccaa
     4981 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5041 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tgcgaaagcc
     5101 tcctgctgat cctgtctctt gatagatgtt tgcaaaagct aggactcgat tatacgttga
     5161 ccgatcagac tatcaatatc taggtttaaa cagatcggta cgctgccgtg accaggcttg
     5221 agtacaatat ctgacgattg gacttcctgg gtcccaagct ggattccgac tggctgactg
     5281 ccaagcaacg atatggatcg tcgagactcg cactgagagg atcgttctcg gtacaccagg
     5341 cgcgcctagc gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt
     5401 ttattatcta agactaggct tacatactag cttattcctt tgccctcgga cgagtgctgg
     5461 ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag acggcggcgc
     5521 ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg attgcgtcgc
     5581 atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc tgatagagtt
     5641 ggtcaagacc aatgcggagc atatacgccc ggagccgggg cgatcctgca agctccggat
     5701 gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc ctccagaaga
     5761 agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag tcaatgaccg
     5821 ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg tgcacgaggt
     5881 gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc tgcgcgacgg
     5941 acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga tcagcaatcg
     6001 cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg aatgggccga
     6061 acccgctcgt ctggctaaga tcggcggcag cgatggcatc cattgcctcc gcgaccggct
     6121 gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc tgtgcacggc
     6181 gggagatgca ataggtcagg ctctcgctga actccccaat gtcaagcact tccggaatcg
     6241 ggagcgcggc ggatgcaaag tgccgataaa cataacgatc tttgtagaaa ccatcggcgc
     6301 agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa gcacgagatt
     6361 cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg atcagaaact
     6421 tctcgacaga cgtcgcggtg agttcaggct ttttacccat gttgtctaac aaaaaagcca
     6481 aaaacggcca gaatttagcg gacaatttac aagtctaaca ctgaaaatta catattgacc
     6541 caaatgatta catttcaaaa ggtgcctaaa aaacttcaca aaacacactc gccaaccccg
     6601 agcgcatagt tcaaaaccgg agcttcagct acttaagaag ataggtacat aaaaccgacc
     6661 aaagaaactg acgcctcact tatccctccc ctcaccagag gtccggcgcc tgtcgattca
     6721 ggagagccta cccta
//
