LOCUS       sequence-300592-        6635 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 6635)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6635)
  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-m4252871
FEATURES             Location/Qualifiers
     source          1..6635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      53..99
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    104..175
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(327..463)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(476..1267,1268..1336))
                     /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     1099..1118
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1602..1650
                     /note="synthetic polyadenylation signal"
     CDS             complement(1677..2276)
                     /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(1760..1780)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2257..2276
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2280..3489)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2424..2443)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      3989..4577
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4478..4497
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4803..4874
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4919..5055)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5085..5131)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5246..5367
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5283..5302)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5337..5356
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5444..5461)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(5810..5830)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(6034..6236)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6063..6083)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6238..6541
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    join(6622..6635,1..35)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgcaacgatt gatagcataa
       61 ccccttgggg cctctaaacg ggtcttgagg ggttttttga attgagaggt agctgaagct
      121 gctgttatga ccactagagg gaagaagata cctgtggagc taatggtcca agatgtcggg
      181 agccgcgatc gctgccaaaa aggtaagctg acgatagctg acgagattcg atgcccgacg
      241 agcaaatgag cgccacggct gttcgcgtaa gctcgaaacg acggagattc gatagagcca
      301 gagtcgagtt taaacgattc gggatcttaa ggataaattt ctggtaagga ggacacgtat
      361 ggaagtgggc aagttgggga agccgtatcc gttgctgaat ctggcatatg tgggagtata
      421 agacgcgcag cgtcgcatca ggcatttttt tctgcgccaa tgcacctaga atcggttacc
      481 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
      541 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
      601 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
      661 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
      721 tcaactgttg ccgggaagct agagtaagta gttcgccagt taatagttta cgcaaggttg
      781 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
      841 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
      901 gctccttcgg tcctccaatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
      961 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     1021 ctggtgaata ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     1081 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     1141 ttggaaagcg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     1201 cgatgtaacc cactcgcgca cccaactgat cttcagcatc ttttactttc accagcgttt
     1261 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     1321 aatgttgaat actcatgcga aagcctcctg ctgatcctgt ctcttgatag atgtttgcaa
     1381 aagctaggac tcgattatac gttgaccgat cagactatca atatctaggt ttaaacagat
     1441 cggtacgctg ccgtgaccag gcttgagtac aatatctgac gattggactt cctgggtccc
     1501 aagctggatt ccgactggct gactgccaag caacgatatg gatcgtcgag actcgcactg
     1561 agaggatcgt tctcggtaca ccaggcgcgc ctagcgttac acacacaaaa aaccaacaca
     1621 catccatctt cgatggatag cgattttatt atctaagact aggcttacat actagcctag
     1681 gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg
     1741 gtgacggtga agccgagccg ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg
     1801 aaggcgggca ccccggcgcg ctcggcggcc tccactccgg ggagcacgac ggcgctgccc
     1861 agacccttgc cctggtggtc gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc
     1921 ttgggccggt gcggcgccag gaggccctcc atctgttgct gcgcggccag ccgggaaccg
     1981 ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc
     2041 ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg
     2101 agcccgacgc gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc
     2161 ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg
     2221 gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatgttg
     2281 tctaacaaaa aagccaaaaa cggccagaat ttagcggaca atttacaagt ctaacactga
     2341 aaattacata ttgacccaaa tgattacatt tcaaaaggtg cctaaaaaac ttcacaaaac
     2401 acactcgcca accccgagcg catagttcaa aaccggagct tcagctactt aagaagatag
     2461 gtacataaaa ccgaccaaag aaactgacgc ctcacttatc cctcccctca ccagaggtcc
     2521 ggcgcctgtc gattcaggag agcctaccct aggcccgaac cctgcgtcct gcgacggaga
     2581 aaagcctacc gcacacctac cggctggtgg ccccaccctg cattataagc caacagaacg
     2641 ggtgacgtca cgacacgacg agggcgcgcg ctcccaaagg tacgggtgca ctgcccaacg
     2701 gcaccgccat aactgccgcc cccgcaacag acgacaaacc gagttctcca gtcagtgaca
     2761 aacttcacgt cagggtcccc agatggtgcc cagcccatct cacccgaata agagctttcc
     2821 cgcattagcg aagggctcaa gaccttgggt tcttgccgcc caccatgccc cccaccttgt
     2881 ttcaacgacc tcacagcccg cctcacaagc gtcttccatt caagactcgg gaacagccgc
     2941 cattttgctg cgctcccccc aacccccagt tcagggcaac cttgctcgcg gacccagact
     3001 acagcccttg gcggtctctc cacacgcttc cgtcccaccg agcggcccgg cggccacgaa
     3061 agccccggcc agcccagcag cccgctactc accaagtgac gatcacagcg atccacaaac
     3121 aagaaccgcg acccaaatcc cggctgcgac ggaactagct gtgccacacc cggcgcgtcc
     3181 ttatataatc atcggcgttc accgccccac ggagatccct ccgcagaatc gccgagaagg
     3241 gactactttt cctcgcctgt tccgctctct ggaaagaaaa ccagtgccct agagtcaccc
     3301 aagtcccgtc ctaaaatgtc cttctgctga tactggggtt ctaaggccga gtcttatgag
     3361 cagcgggccg ctgtcctgag cgtccgggcg gaaggatcag gacgctcctg cgcccttcgt
     3421 ctgacgtggc agcgctcgcc gtgacgaggg gggcgcccgc gggaggcgcc aaaacccggc
     3481 gcggaggcca ggcgcgcccc gatttcgata ttcctcggat ctcggataga tcgggattga
     3541 ggatgtaggt tgaacttctc tcgggtgatc ccgatctcgg atattcggga ttggagtaat
     3601 acctggcgat aggcgctgcc aagcaatagc caaggcgcaa ggccggccaa gagacttgcg
     3661 atagttcgtt cgagaggtag cctgcaagat ggatcggacg aaaggtttga agattttccg
     3721 acgagggcct tggagagttc ggaaatttaa atgcaactat ggatgaacga aatagacaga
     3781 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3841 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3901 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3961 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4021 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4081 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4141 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4201 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4261 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4321 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4381 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4441 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4501 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4561 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4621 ggccttttgc tcacatgatt taaatgaaac tcgtaatcgg agaacgggtg caatagcaag
     4681 gaccagatcg aacttgactg acgcaattct gagcgtagtc gaaacccaga caagtcgatc
     4741 tcgccaagta gctgacttga ctggttaatt aagtttcgat agcccaaggt aaccaatgag
     4801 attagggagg gaagaaagtt tgatgaacta cttctgaccc ctagtggtgt ccagaaaaga
     4861 ccattaaagg aatgtcttga tcgaaatcga gagatcgcct gcaggatagc tgacgacctt
     4921 aaggataaat ttctggtaag gaggacacgt atggaagtgg gcaagttggg gaagccgtat
     4981 ccgttgctga atctggcata tgtgggagta taagacgcgc agcgtcgcat caggcatttt
     5041 tttctgcgcc aatgccggta gatagcaatc cggcccgagg ggcacaaaaa acccctcaag
     5101 acccgtttag aggccccaag gggttatgct acgataccga tagagatggg ccaataccac
     5161 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     5221 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     5281 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5341 tttgtccaaa ctcatcaatg tatcttatca tgtctgtgag ctgaaggtac gctgtatctc
     5401 agtcagtcaa gctagcttag ccgcgtggca cgagcccgct gctatggtga tggtgatggt
     5461 gcatgtgccc cgagcctgaa cctgccaggt tagcgtccaa gaactctttt agctggccct
     5521 tagacagggc tcccactttt gtcgcggcaa cttccccatt cttaaagagc aggagggtag
     5581 gtattcccct gatgccgtat ttaggtgccg tccctgggtt ctgatctata ttaagcttag
     5641 cgacagttaa ttttccttga tactcgtcgg cgatctcgtc cagaatgggt gcgatcatct
     5701 tgcacgggcc acaccattca gcccagaaat ctaccagtat cgcaccatcg gccttcaaca
     5761 cgtctgtatc aaaggagtca tcggttagat gaatgatttt gtcactcatc gactgaaagt
     5821 aaaggttttc ggcgtgacat gtctcattat gatcttgtaa ctccagactg aaccaaaaaa
     5881 tgattagcat aatgagcacc agaaaaagca ggaaagccaa gaagcacaga taaaagtcga
     5941 tgagcgatag ttctatcatg gtggcggccg ctggcgggtg tgtcgagtgg atggtaggat
     6001 cgacaaagat ctgatctgac ggttcactaa accagctctg cttatataga cctcccaccg
     6061 tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc
     6121 ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa
     6181 tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcagcat
     6241 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat
     6301 gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact
     6361 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca
     6421 ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt
     6481 caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac
     6541 gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact agatctggct
     6601 acactcgatc agcagttaga taataaaatc gctat
//
