LOCUS       sequence-307328-        6708 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 6708)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6708)
  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-m4252957
FEATURES             Location/Qualifiers
     source          1..6708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    21..69
                     /note="synthetic polyadenylation signal"
     terminator      87..133
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    138..209
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(361..497)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(510..1301,1302..1370))
                     /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     1133..1152
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1636..1684
                     /note="synthetic polyadenylation signal"
     CDS             complement(1711..2739)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2743..3952)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2887..2906)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4452..5040
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4941..4960
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5266..5337
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5382..5518)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5548..5594)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5709..5830
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5746..5765)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5800..5819
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5899..5916)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(5917..5937)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(6141..6343)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6170..6190)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6345..6648
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cactcgatca gcagttagat aataaaatcg ctatccatcg aagatggatg tgtgttggtt
       61 ttttgtgtgt gtaacgcaac gattgatagc ataacccctt ggggcctcta aacgggtctt
      121 gaggggtttt ttgaattgag aggtagctga agctgctgtt atgaccacta gagggaagaa
      181 gatacctgtg gagctaatgg tccaagatgt cgggagccgc gatcgctgcc aaaaaggtaa
      241 gctgacgata gctgacgaga ttcgatgccc gacgagcaaa tgagcgccac ggctgttcgc
      301 gtaagctcga aacgacggag attcgataga gccagagtcg agtttaaacg attcgggatc
      361 ttaaggataa atttctggta aggaggacac gtatggaagt gggcaagttg gggaagccgt
      421 atccgttgct gaatctggca tatgtgggag tataagacgc gcagcgtcgc atcaggcatt
      481 tttttctgcg ccaatgcacc tagaatcggt taccaatgct taatcagtga ggcacctatc
      541 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
      601 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
      661 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
      721 ggtcctgcaa ctttatccgc ctccatccag tctatcaact gttgccggga agctagagta
      781 agtagttcgc cagttaatag tttacgcaag gttgttgcca ttgctacagg catcgtggtg
      841 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
      901 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc aatcgttgtc
      961 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1021 actgtcatgc catccgtaag atgcttttct gtgactggtg aatactcaac caagtcattc
     1081 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1141 gcgccacata gcagaacttt aaaagtgctc atcattggaa agcgttcttc ggggcgaaaa
     1201 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg cgcacccaac
     1261 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1321 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat gcgaaagcct
     1381 cctgctgatc ctgtctcttg atagatgttt gcaaaagcta ggactcgatt atacgttgac
     1441 cgatcagact atcaatatct aggtttaaac agatcggtac gctgccgtga ccaggcttga
     1501 gtacaatatc tgacgattgg acttcctggg tcccaagctg gattccgact ggctgactgc
     1561 caagcaacga tatggatcgt cgagactcgc actgagagga tcgttctcgg tacaccaggc
     1621 gcgcctagcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg atagcgattt
     1681 tattatctaa gactaggctt acatactagc ttattccttt gccctcggac gagtgctggg
     1741 gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga cggcggcgct
     1801 tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga ttgcgtcgca
     1861 tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct gatagagttg
     1921 gtcaagacca atgcggagca tatacgcccg gagccggggc gatcctgcaa gctccggatg
     1981 cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc tccagaagaa
     2041 gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt caatgaccgc
     2101 tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt gcacgaggtg
     2161 ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct gcgcgacgga
     2221 cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat cagcaatcgc
     2281 gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga atgggccgaa
     2341 cccgctcgtc tggctaagat cggcggcagc gatggcatcc attgcctccg cgaccggctg
     2401 cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg
     2461 ggagatgcaa taggtcaggc tctcgctgaa ctccccaatg tcaagcactt ccggaatcgg
     2521 gagcgcggcg gatgcaaagt gccgataaac ataacgatct ttgtagaaac catcggcgca
     2581 gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag cacgagattc
     2641 ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga tcagaaactt
     2701 ctcgacagac gtcgcggtga gttcaggctt tttacccatg ttgtctaaca aaaaagccaa
     2761 aaacggccag aatttagcgg acaatttaca agtctaacac tgaaaattac atattgaccc
     2821 aaatgattac atttcaaaag gtgcctaaaa aacttcacaa aacacactcg ccaaccccga
     2881 gcgcatagtt caaaaccgga gcttcagcta cttaagaaga taggtacata aaaccgacca
     2941 aagaaactga cgcctcactt atccctcccc tcaccagagg tccggcgcct gtcgattcag
     3001 gagagcctac cctaggcccg aaccctgcgt cctgcgacgg agaaaagcct accgcacacc
     3061 taccggctgg tggccccacc ctgcattata agccaacaga acgggtgacg tcacgacacg
     3121 acgagggcgc gcgctcccaa aggtacgggt gcactgccca acggcaccgc cataactgcc
     3181 gcccccgcaa cagacgacaa accgagttct ccagtcagtg acaaacttca cgtcagggtc
     3241 cccagatggt gcccagccca tctcacccga ataagagctt tcccgcatta gcgaagggct
     3301 caagaccttg ggttcttgcc gcccaccatg ccccccacct tgtttcaacg acctcacagc
     3361 ccgcctcaca agcgtcttcc attcaagact cgggaacagc cgccattttg ctgcgctccc
     3421 cccaaccccc agttcagggc aaccttgctc gcggacccag actacagccc ttggcggtct
     3481 ctccacacgc ttccgtccca ccgagcggcc cggcggccac gaaagccccg gccagcccag
     3541 cagcccgcta ctcaccaagt gacgatcaca gcgatccaca aacaagaacc gcgacccaaa
     3601 tcccggctgc gacggaacta gctgtgccac acccggcgcg tccttatata atcatcggcg
     3661 ttcaccgccc cacggagatc cctccgcaga atcgccgaga agggactact tttcctcgcc
     3721 tgttccgctc tctggaaaga aaaccagtgc cctagagtca cccaagtccc gtcctaaaat
     3781 gtccttctgc tgatactggg gttctaaggc cgagtcttat gagcagcggg ccgctgtcct
     3841 gagcgtccgg gcggaaggat caggacgctc ctgcgccctt cgtctgacgt ggcagcgctc
     3901 gccgtgacga ggggggcgcc cgcgggaggc gccaaaaccc ggcgcggagg ccaggcgcgc
     3961 cccgatttcg atattcctcg gatctcggat agatcgggat tgaggatgta ggttgaactt
     4021 ctctcgggtg atcccgatct cggatattcg ggattggagt aatacctggc gataggcgct
     4081 gccaagcaat agccaaggcg caaggccggc caagagactt gcgatagttc gttcgagagg
     4141 tagcctgcaa gatggatcgg acgaaaggtt tgaagatttt ccgacgaggg ccttggagag
     4201 ttcggaaatt taaatgcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4261 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4321 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4381 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4441 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4501 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4561 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4621 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4681 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4741 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4801 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4861 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4921 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4981 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5041 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5101 atttaaatga aactcgtaat cggagaacgg gtgcaatagc aaggaccaga tcgaacttga
     5161 ctgacgcaat tctgagcgta gtcgaaaccc agacaagtcg atctcgccaa gtagctgact
     5221 tgactggtta attaagtttc gatagcccaa ggtaaccaat gagattaggg agggaagaaa
     5281 gtttgatgaa ctacttctga cccctagtgg tgtccagaaa agaccattaa aggaatgtct
     5341 tgatcgaaat cgagagatcg cctgcaggat agctgacgac cttaaggata aatttctggt
     5401 aaggaggaca cgtatggaag tgggcaagtt ggggaagccg tatccgttgc tgaatctggc
     5461 atatgtggga gtataagacg cgcagcgtcg catcaggcat ttttttctgc gccaatgccg
     5521 gtagatagca atccggcccg aggggcacaa aaaacccctc aagacccgtt tagaggcccc
     5581 aaggggttat gctacgatac cgatagagat gggccaatac cacatttgta gaggttttac
     5641 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     5701 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5761 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5821 atgtatctta tcatgtctgt gagctgaagg tacgctgtat ctcagtcagt caagctagca
     5881 gaggcctgcg gatccttaat ggtgatggtg atggtgcgac tgaaagtaaa ggttttcggc
     5941 gtgacaagtt tcattatgat cttgcagttc aagtgagaac caaaagataa taagcataat
     6001 taaaacaagg aatagcagaa aggctaaaaa gcacaaatag aagtcaatta atgaaagttc
     6061 aatcatggtg gcggccgctg gcgggtgtgt cgagtggatg gtaggatcga caaagatctg
     6121 atctgacggt tcactaaacc agctctgctt atatagacct cccaccgtac acgcctaccg
     6181 cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg
     6241 tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca
     6301 aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcagcatggt aatagcgatg
     6361 actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa
     6421 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac
     6481 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg
     6541 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg
     6601 tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata
     6661 tatgggctat gaactaatga ccccgtaatt gattactaga tctggcta
//
