LOCUS       sequence-358357-        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"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             712..732
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             733..774
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    843..964
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(854..873)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     908..927
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1079..1125
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1155..1291
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1336..1407
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1633..2221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1713..1732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2721..3930
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3767..3786
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3934..4962
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4989..5037
                     /note="synthetic polyadenylation signal"
     CDS             join(5303..5371,5372..6163)
                     /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(5521..5540)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6176..6312
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6464..6535
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6540..6586)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6604..6652
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgattga actttcatta
      601 attgacttct atttgtgctt tttagccttt ctgctattcc ttgttttaat tatgcttatt
      661 atcttttggt tctcacttga actgcaagat cataatgaaa cttgtcacgc cgaaaacctt
      721 tactttcagt cggggaaacc tatacccaat ccacttctcg gcttagattc tacctaagga
      781 tccgcaggcc tctgctagct tgactgactg agatacagcg taccttcagc tcacagacat
      841 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
      901 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
      961 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1021 tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta
     1081 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg
     1141 attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     1201 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     1261 cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct
     1321 cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt
     1381 agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta
     1441 attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca
     1501 gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag
     1561 tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1621 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1681 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1741 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1801 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1861 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1921 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1981 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2041 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2101 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2161 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2221 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2281 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2341 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2401 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt
     2461 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca
     2521 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg
     2581 ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg
     2641 atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat
     2701 atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc
     2761 cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc aggagcgtcc tgatccttcc
     2821 gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga accccagtat
     2881 cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt tttctttcca
     2941 gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga gggatctccg
     3001 tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca gctagttccg
     3061 tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg tcacttggtg
     3121 agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc ggtgggacgg
     3181 aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa
     3241 ctgggggttg gggggagcgc agcaaaatgg cggctgttcc cgagtcttga atggaagacg
     3301 cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa
     3361 cccaaggtct tgagcccttc gctaatgcgg gaaagctctt attcgggtga gatgggctgg
     3421 gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc
     3481 tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac ctttgggagc
     3541 gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc agggtggggc
     3601 caccagccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta
     3661 gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg ataagtgagg
     3721 cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa gctccggttt
     3781 tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa
     3841 atgtaatcat ttgggtcaat atgtaatttt cagtgttaga cttgtaaatt gtccgctaaa
     3901 ttctggccgt ttttggcttt tttgttagac aacatgggta aaaagcctga actcaccgcg
     3961 acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc
     4021 tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg
     4081 cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca
     4141 tccgccgcgc tcccgattcc ggaagtgctt gacattgggg agttcagcga gagcctgacc
     4201 tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg
     4261 cccgctgttc tgcagccggt cgcggaggca atggatgcca tcgctgccgc cgatcttagc
     4321 cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt
     4381 gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac
     4441 accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc
     4501 cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat
     4561 ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag
     4621 gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac
     4681 ttcgagcgga ggcatccgga gcttgcagga tcgccccggc tccgggcgta tatgctccgc
     4741 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg
     4801 gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa
     4861 atcgcccgca gaagcgccgc cgtctggacc gatggctgtg tagaagtact cgccgatagt
     4921 ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat aagctagtat gtaagcctag
     4981 tcttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt
     5041 aacgctaggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca
     5101 tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt
     5161 cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg
     5221 atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac
     5281 aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5341 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5401 tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa
     5461 gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct
     5521 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5581 acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga
     5641 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5701 caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat
     5761 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5821 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac
     5881 tggcgaacta cttactctag cttcccggca acagttgata gactggatgg aggcggataa
     5941 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6001 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6061 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6121 acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt
     6181 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     6241 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     6301 atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg
     6361 tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca
     6421 gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc
     6481 tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc
     6541 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt
     6601 acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact
     6661 gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata
     6721 tatggagttc cg
//
