LOCUS       sequence-300755-        6717 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 6717)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6717)
  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-m4252966
FEATURES             Location/Qualifiers
     source          1..6717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    74..145
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(150..196)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    214..262
                     /note="synthetic polyadenylation signal"
     enhancer        343..646
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        648..850
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     801..821
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1054..1074
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1075..1101
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1170..1291
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1181..1200)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1235..1254
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1406..1452
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1482..1618
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1663..1734
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1960..2548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2040..2059)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3048..4257
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4094..4113
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4261..5289
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5316..5364
                     /note="synthetic polyadenylation signal"
     CDS             join(5630..5698,5699..6490)
                     /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(5848..5867)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6503..6639
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga
       61 tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc
      121 ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc
      181 ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc
      241 ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc
      301 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      361 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      421 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      481 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      541 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      601 tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg
      661 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      721 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      781 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      841 aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg
      901 acacacccgc cagcggccgc caccatgatt gaactttcat taattgactt ctatttgtgc
      961 tttttagcct ttctgctatt ccttgtttta attatgctta ttatcttttg gttctcactt
     1021 gaactgcaag atcataatga aacttgtcac gccgaaaacc tttactttca gtcgtacccc
     1081 tatgacgtac cagattatgc gtaaggatcc gcaggcctct gctagcttga ctgactgaga
     1141 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
     1201 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1261 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1321 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     1381 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
     1441 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
     1501 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     1561 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
     1621 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
     1681 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
     1741 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
     1801 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
     1861 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     1921 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1981 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2041 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2101 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2161 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2221 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2281 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2341 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2401 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2461 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2521 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2581 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2641 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2701 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2761 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     2821 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     2881 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     2941 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     3001 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg
     3061 ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca cgtcagacga
     3121 agggcgcagg agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca
     3181 taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt
     3241 gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt
     3301 ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac
     3361 gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt
     3421 gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg
     3481 tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc
     3541 tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg
     3601 ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa
     3661 ggtggggggc atggtgggcg gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa
     3721 agctcttatt cgggtgagat gggctgggca ccatctgggg accctgacgt gaagtttgtc
     3781 actgactgga gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt
     3841 gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt
     3901 ctgttggctt ataatgcagg gtggggccac cagccggtag gtgtgcggta ggcttttctc
     3961 cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac
     4021 ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat
     4081 cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt
     4141 tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag
     4201 tgttagactt gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacaac
     4261 atgggtaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     4321 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     4381 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     4441 gatcgttatg tttatcggca ctttgcatcc gccgcgctcc cgattccgga agtgcttgac
     4501 attggggagt tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
     4561 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggcaatg
     4621 gatgccatcg ctgccgccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     4681 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     4741 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     4801 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     4861 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     4921 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     4981 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     5041 ccccggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     5101 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     5161 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgccgccgt ctggaccgat
     5221 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     5281 aaggaataag ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat
     5341 ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat
     5401 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     5461 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     5521 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     5581 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     5641 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5701 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     5761 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     5821 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5881 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     5941 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6001 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     6061 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6121 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6181 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6241 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6301 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6361 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6421 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6481 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     6541 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     6601 cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc
     6661 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgct
//
