LOCUS       sequence-297326-        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"
     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    268..389
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(279..298)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     333..352
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      504..550
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      580..716
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    761..832
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1058..1646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1138..1157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2146..3355
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3192..3211
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3359..4387
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4414..4462
                     /note="synthetic polyadenylation signal"
     CDS             join(4728..4796,4797..5588)
                     /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(4946..4965)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5601..5737
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5889..5960
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5965..6011)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6029..6077
                     /note="synthetic polyadenylation signal"
     enhancer        6158..6461
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6463..6665
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6616..6636
                     /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 aaggatccgc aggcctctgc tagcttgact gactgagata
      241 cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa
      301 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
      361 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
      421 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
      481 ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag
      541 gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg
      601 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
      661 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc
      721 gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt
      781 ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt
      841 ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga
      901 cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct
      961 attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa
     1021 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1081 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1141 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1201 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1261 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1321 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1381 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1441 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     1501 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1561 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1621 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1681 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1741 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1801 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1861 tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa
     1921 atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag
     1981 tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact
     2041 ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc
     2101 ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggcct ccgcgccggg
     2161 ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag cgctgccacg tcagacgaag
     2221 ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata
     2281 agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga
     2341 ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct
     2401 cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc
     2461 gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt
     2521 ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctggccgg ggctttcgtg
     2581 gccgccgggc cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg
     2641 ggtccgcgag caaggttgcc ctgaactggg ggttgggggg agcgcagcaa aatggcggct
     2701 gttcccgagt cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg
     2761 tggggggcat ggtgggcggc aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag
     2821 ctcttattcg ggtgagatgg gctgggcacc atctggggac cctgacgtga agtttgtcac
     2881 tgactggaga actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg
     2941 gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct
     3001 gttggcttat aatgcagggt ggggccacca gccggtaggt gtgcggtagg cttttctccg
     3061 tcgcaggacg cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct
     3121 ctggtgaggg gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct
     3181 tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg
     3241 tgaagttttt taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg
     3301 ttagacttgt aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacaacat
     3361 gggtaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     3421 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     3481 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     3541 tcgttatgtt tatcggcact ttgcatccgc cgcgctcccg attccggaag tgcttgacat
     3601 tggggagttc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     3661 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggcaatgga
     3721 tgccatcgct gccgccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     3781 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     3841 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     3901 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     3961 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     4021 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     4081 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     4141 ccggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     4201 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     4261 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gccgccgtct ggaccgatgg
     4321 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     4381 ggaataagct agtatgtaag cctagtctta gataataaaa tcgctatcca tcgaagatgg
     4441 atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc
     4501 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     4561 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     4621 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     4681 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     4741 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4801 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     4861 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     4921 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4981 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     5041 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5101 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     5161 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5221 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5281 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     5341 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5401 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5461 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5521 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5581 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     5641 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     5701 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga
     5761 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     5821 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     5881 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     5941 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     6001 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga
     6061 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta
     6121 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     6181 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     6241 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     6301 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     6361 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     6421 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt
     6481 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     6541 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     6601 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     6661 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca
     6721 cccgccagcg gccgc
//
