LOCUS       sequence-300688-        7245 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 7245)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7245)
  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-m4252909
FEATURES             Location/Qualifiers
     source          1..7245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    74..122
                     /note="synthetic polyadenylation signal"
     terminator      140..186
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    191..262
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(414..550)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(563..1354,1355..1423))
                     /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     1186..1205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1689..1737
                     /note="synthetic polyadenylation signal"
     CDS             complement(1764..2792)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2796..4005)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2940..2959)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4505..5093
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4994..5013
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5319..5390
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5435..5571)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5601..5647)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5762..5883
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5799..5818)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5853..5872
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(6348..6368)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(6396..6473)
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             complement(6507..6527)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(6731..6933)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6760..6780)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6935..7238
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ccatatatgg gctatgaact aatgaccccg taattgatta ctagatctgg ctacactcga
       61 tcagcagtta gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg
      121 tgtgtaacgc aacgattgat agcataaccc cttggggcct ctaaacgggt cttgaggggt
      181 tttttgaatt gagaggtagc tgaagctgct gttatgacca ctagagggaa gaagatacct
      241 gtggagctaa tggtccaaga tgtcgggagc cgcgatcgct gccaaaaagg taagctgacg
      301 atagctgacg agattcgatg cccgacgagc aaatgagcgc cacggctgtt cgcgtaagct
      361 cgaaacgacg gagattcgat agagccagag tcgagtttaa acgattcggg atcttaagga
      421 taaatttctg gtaaggagga cacgtatgga agtgggcaag ttggggaagc cgtatccgtt
      481 gctgaatctg gcatatgtgg gagtataaga cgcgcagcgt cgcatcaggc atttttttct
      541 gcgccaatgc acctagaatc ggttaccaat gcttaatcag tgaggcacct atctcagcga
      601 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
      661 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
      721 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
      781 caactttatc cgcctccatc cagtctatca actgttgccg ggaagctaga gtaagtagtt
      841 cgccagttaa tagtttacgc aaggttgttg ccattgctac aggcatcgtg gtgtcacgct
      901 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
      961 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccaatcgtt gtcagaagta
     1021 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     1081 tgccatccgt aagatgcttt tctgtgactg gtgaatactc aaccaagtca ttctgagaat
     1141 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     1201 atagcagaac tttaaaagtg ctcatcattg gaaagcgttc ttcggggcga aaactctcaa
     1261 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgcgcaccc aactgatctt
     1321 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     1381 caaaaaaggg aataagggcg acacggaaat gttgaatact catgcgaaag cctcctgctg
     1441 atcctgtctc ttgatagatg tttgcaaaag ctaggactcg attatacgtt gaccgatcag
     1501 actatcaata tctaggttta aacagatcgg tacgctgccg tgaccaggct tgagtacaat
     1561 atctgacgat tggacttcct gggtcccaag ctggattccg actggctgac tgccaagcaa
     1621 cgatatggat cgtcgagact cgcactgaga ggatcgttct cggtacacca ggcgcgccta
     1681 gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc
     1741 taagactagg cttacatact agcttattcc tttgccctcg gacgagtgct ggggcgtcgg
     1801 tttccactat cggcgagtac ttctacacag ccatcggtcc agacggcggc gcttctgcgg
     1861 gcgatttgtg tacgcccgac agtcccggct ccggatcgga cgattgcgtc gcatcgaccc
     1921 tgcgcccaag ctgcatcatc gaaattgccg tcaaccaagc tctgatagag ttggtcaaga
     1981 ccaatgcgga gcatatacgc ccggagccgg ggcgatcctg caagctccgg atgcctccgc
     2041 tcgaagtagc gcgtctgctg ctccatacaa gccaaccacg gcctccagaa gaagatgttg
     2101 gcgacctcgt attgggaatc cccgaacatc gcctcgctcc agtcaatgac cgctgttatg
     2161 cggccattgt ccgtcaggac attgttggag ccgaaatccg cgtgcacgag gtgccggact
     2221 tcggggcagt cctcggccca aagcatcagc tcatcgagag cctgcgcgac ggacgcactg
     2281 acggtgtcgt ccatcacagt ttgccagtga tacacatggg gatcagcaat cgcgcatatg
     2341 aaatcacgcc atgtagtgta ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc
     2401 gtctggctaa gatcggcggc agcgatggca tccattgcct ccgcgaccgg ctgcagaaca
     2461 gcgggcagtt cggtttcagg caggtcttgc aacgtgacac cctgtgcacg gcgggagatg
     2521 caataggtca ggctctcgct gaactcccca atgtcaagca cttccggaat cgggagcgcg
     2581 gcggatgcaa agtgccgata aacataacga tctttgtaga aaccatcggc gcagctattt
     2641 acccgcagga catatccacg ccctcctaca tcgaagctga aagcacgaga ttcttcgccc
     2701 tccgagagct gcatcaggtc ggagacgctg tcgaactttt cgatcagaaa cttctcgaca
     2761 gacgtcgcgg tgagttcagg ctttttaccc atgttgtcta acaaaaaagc caaaaacggc
     2821 cagaatttag cggacaattt acaagtctaa cactgaaaat tacatattga cccaaatgat
     2881 tacatttcaa aaggtgccta aaaaacttca caaaacacac tcgccaaccc cgagcgcata
     2941 gttcaaaacc ggagcttcag ctacttaaga agataggtac ataaaaccga ccaaagaaac
     3001 tgacgcctca cttatccctc ccctcaccag aggtccggcg cctgtcgatt caggagagcc
     3061 taccctaggc ccgaaccctg cgtcctgcga cggagaaaag cctaccgcac acctaccggc
     3121 tggtggcccc accctgcatt ataagccaac agaacgggtg acgtcacgac acgacgaggg
     3181 cgcgcgctcc caaaggtacg ggtgcactgc ccaacggcac cgccataact gccgcccccg
     3241 caacagacga caaaccgagt tctccagtca gtgacaaact tcacgtcagg gtccccagat
     3301 ggtgcccagc ccatctcacc cgaataagag ctttcccgca ttagcgaagg gctcaagacc
     3361 ttgggttctt gccgcccacc atgcccccca ccttgtttca acgacctcac agcccgcctc
     3421 acaagcgtct tccattcaag actcgggaac agccgccatt ttgctgcgct ccccccaacc
     3481 cccagttcag ggcaaccttg ctcgcggacc cagactacag cccttggcgg tctctccaca
     3541 cgcttccgtc ccaccgagcg gcccggcggc cacgaaagcc ccggccagcc cagcagcccg
     3601 ctactcacca agtgacgatc acagcgatcc acaaacaaga accgcgaccc aaatcccggc
     3661 tgcgacggaa ctagctgtgc cacacccggc gcgtccttat ataatcatcg gcgttcaccg
     3721 ccccacggag atccctccgc agaatcgccg agaagggact acttttcctc gcctgttccg
     3781 ctctctggaa agaaaaccag tgccctagag tcacccaagt cccgtcctaa aatgtccttc
     3841 tgctgatact ggggttctaa ggccgagtct tatgagcagc gggccgctgt cctgagcgtc
     3901 cgggcggaag gatcaggacg ctcctgcgcc cttcgtctga cgtggcagcg ctcgccgtga
     3961 cgaggggggc gcccgcggga ggcgccaaaa cccggcgcgg aggccaggcg cgccccgatt
     4021 tcgatattcc tcggatctcg gatagatcgg gattgaggat gtaggttgaa cttctctcgg
     4081 gtgatcccga tctcggatat tcgggattgg agtaatacct ggcgataggc gctgccaagc
     4141 aatagccaag gcgcaaggcc ggccaagaga cttgcgatag ttcgttcgag aggtagcctg
     4201 caagatggat cggacgaaag gtttgaagat tttccgacga gggccttgga gagttcggaa
     4261 atttaaatgc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4321 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4381 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4441 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4501 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4561 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4621 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4681 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4741 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4801 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4861 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4921 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4981 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5041 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5101 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgatttaaa
     5161 tgaaactcgt aatcggagaa cgggtgcaat agcaaggacc agatcgaact tgactgacgc
     5221 aattctgagc gtagtcgaaa cccagacaag tcgatctcgc caagtagctg acttgactgg
     5281 ttaattaagt ttcgatagcc caaggtaacc aatgagatta gggagggaag aaagtttgat
     5341 gaactacttc tgacccctag tggtgtccag aaaagaccat taaaggaatg tcttgatcga
     5401 aatcgagaga tcgcctgcag gatagctgac gaccttaagg ataaatttct ggtaaggagg
     5461 acacgtatgg aagtgggcaa gttggggaag ccgtatccgt tgctgaatct ggcatatgtg
     5521 ggagtataag acgcgcagcg tcgcatcagg catttttttc tgcgccaatg ccggtagata
     5581 gcaatccggc ccgaggggca caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     5641 tatgctacga taccgataga gatgggccaa taccacattt gtagaggttt tacttgcttt
     5701 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     5761 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5821 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5881 ttatcatgtc tgtgagctga aggtacgctg tatctcagtc agtcaagcta gcagaggcct
     5941 gcggatcctt attgatggtt cgcgtctact ttcggggcct gggcatcatt gagctttttg
     6001 gcttccgcaa gcaggttggc actctgtgat ggatcgtctt ttagggactg gataaatgca
     6061 tttctttgtt cctcattcag attaggaagg tgcaatatct cgtagaaagc attctgctgc
     6121 tccttgttaa acttgttatc aacttttgga gcctgtgcgt cattcagctt cttggcttcg
     6181 gccaggagat ttgcagactg cgagggatcg tctttcaggc tctgaatgaa agcgttgcgt
     6241 tgctcttcgt tcagatttgg taagtgaagg atctcataga aagcgttctg ctgttcttta
     6301 ttaaacttgt tatcgacggc ctcgtcatgt tgtgccaagc caaattcacc ctggaaatag
     6361 agattctcgg aggcggtcgt aggaatgtcg taatctaggg cgccgcttga actgatcttt
     6421 ttgaaccggt tagccgcaga cactgcaata aagttcttct tccatctcct tttccaggga
     6481 tttatcagcc ccgggattcg ccgtcccgac tgaaagtaaa ggttttcggc gtgacaagtt
     6541 tcattatgat cttgcagttc aagtgagaac caaaagataa taagcataat taaaacaagg
     6601 aatagcagaa aggctaaaaa gcacaaatag aagtcaatta atgaaagttc aatcatggtg
     6661 gcggccgctg gcgggtgtgt cgagtggatg gtaggatcga caaagatctg atctgacggt
     6721 tcactaaacc agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg
     6781 tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca
     6841 aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc
     6901 cacgcccatt gatgtactgc caaaaccgca tcagcatggt aatagcgatg actaatacgt
     6961 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     7021 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     7081 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     7141 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     7201 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaact
//
