LOCUS       sequence-299538-        6732 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 6732)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6732)
  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-m4258315
FEATURES             Location/Qualifiers
     source          1..6732
                     /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..196
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    265..386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(276..295)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     330..349
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      501..547
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      577..713
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    758..829
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1055..1643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1135..1154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2143..3352
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3189..3208
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3356..4384
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4411..4459
                     /note="synthetic polyadenylation signal"
     CDS             join(4725..4793,4794..5585)
                     /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(4943..4962)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5598..5734
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5886..5957
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5962..6008)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6026..6074
                     /note="synthetic polyadenylation signal"
     enhancer        6155..6458
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6460..6662
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6613..6633
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgatt gaactttcat taattgactt ctatttgtgc tttttagcct ttctgctatt
       61 ccttgtttta attatgctta ttatcttttg gttctcactt gaactgcaag atcataatga
      121 aacttgtcac gccgaaaacc tttactttca gtcggggaaa cctataccca atccacttct
      181 cggcttagat tctacctaag gatccgcagg cctctgctag cttgactgac tgagatacag
      241 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
      301 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
      361 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
      421 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
      481 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
      541 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
      601 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
      661 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
      721 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
      781 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
      841 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
      901 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
      961 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
     1021 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1081 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1141 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1201 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1261 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1321 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1381 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1441 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1501 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1561 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1621 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1681 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1741 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1801 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1861 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     1921 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     1981 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     2041 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     2101 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt
     2161 tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc
     2221 gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     2281 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     2341 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     2401 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     2461 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     2521 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     2581 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     2641 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     2701 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     2761 ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc
     2821 ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga
     2881 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca
     2941 gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt
     3001 ggcttataat gcagggtggg gccaccagcc ggtaggtgtg cggtaggctt ttctccgtcg
     3061 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg
     3121 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct
     3181 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga
     3241 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta
     3301 gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatggg
     3361 taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     3421 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     3481 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     3541 ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc ttgacattgg
     3601 ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     3661 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg caatggatgc
     3721 catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     3781 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     3841 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     3901 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     3961 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     4021 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     4081 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccccg
     4141 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     4201 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     4261 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga ccgatggctg
     4321 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     4381 ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg
     4441 tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct
     4501 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     4561 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     4621 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     4681 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     4741 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4801 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     4861 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     4921 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4981 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     5041 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5101 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     5161 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5221 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5281 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     5341 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5401 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5461 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5521 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5581 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     5641 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     5701 atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc
     5761 tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct
     5821 cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct
     5881 cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc
     5941 agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     6001 ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg
     6061 atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg
     6121 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     6181 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     6241 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     6301 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     6361 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     6421 ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca
     6481 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     6541 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     6601 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     6661 ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc
     6721 gccagcggcc gc
//
