LOCUS       sequence-297427-        6287 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 6287)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6287)
  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-m4252833
FEATURES             Location/Qualifiers
     source          1..6287
                     /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    249..370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(260..279)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     314..333
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      485..531
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      561..697
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    742..813
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1039..1627)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1119..1138)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2127..3336
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3173..3192
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3340..3939
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3340..3359)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3836..3856
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3966..4014
                     /note="synthetic polyadenylation signal"
     CDS             join(4280..4348,4349..5140)
                     /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(4498..4517)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5153..5289
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5441..5512
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5517..5563)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5581..5629
                     /note="synthetic polyadenylation signal"
     enhancer        5710..6013
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6015..6217
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6168..6188
                     /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 gtcggggaaa cctataccca atccacttct
      181 cggcttagat tctacctaag ctagcttgac tgactgagat acagcgtacc ttcagctcac
      241 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
      301 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
      361 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
      421 ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt
      481 atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg
      541 ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
      601 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
      661 tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg
      721 atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc
      781 agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga
      841 aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta
      901 cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat
      961 tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1021 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1081 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1141 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1201 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1261 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1321 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1381 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1441 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1501 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1561 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1621 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1681 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1741 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1801 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1861 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc
     1921 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg
     1981 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag
     2041 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg
     2101 aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg
     2161 cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat
     2221 ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc
     2281 cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc
     2341 tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga
     2401 tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta
     2461 gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac
     2521 ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg
     2581 ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc
     2641 cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg
     2701 aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg
     2761 caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc gggtgagatg
     2821 ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt
     2881 gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt
     2941 gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg
     3001 tggggccacc agccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg
     3061 ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa
     3121 gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc
     3181 cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct
     3241 tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg taaattgtcc
     3301 gctaaattct ggccgttttt ggcttttttg ttagacaaca tgaccgagta caagcccacg
     3361 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     3421 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     3481 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     3541 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     3601 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     3661 cagatggagg gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     3721 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     3781 gtggaggccg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     3841 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     3901 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctagg ctagtatgta agcctagtct
     3961 tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac
     4021 gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     4081 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     4141 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     4201 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     4261 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     4321 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4381 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4441 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     4501 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4561 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     4621 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4681 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4741 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4801 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     4861 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     4921 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4981 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5041 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5101 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     5161 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     5221 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     5281 tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     5341 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     5401 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     5461 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     5521 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     5581 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     5641 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     5701 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     5761 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     5821 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     5881 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     5941 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     6001 cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     6061 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     6121 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     6181 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag
     6241 atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
