LOCUS       sequence-300729-        7230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF10 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7230)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7230)
  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-m4252927
FEATURES             Location/Qualifiers
     source          1..7230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      60..106
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    111..182
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(334..470)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(483..1274,1275..1343))
                     /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     1106..1125
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1609..1657
                     /note="synthetic polyadenylation signal"
     CDS             complement(1684..2712)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2716..3925)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2860..2879)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4425..5013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4914..4933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5239..5310
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5355..5491)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5521..5567)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5682..5803
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5719..5738)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5773..5792
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(6268..6288)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(6316..6393)
                     /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(6427..6447)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      6558..6567
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6636..6838)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6665..6685)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6840..7143
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    join(7224..7230,1..42)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc aacgattgat
       61 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgaatt gagaggtagc
      121 tgaagctgct gttatgacca ctagagggaa gaagatacct gtggagctaa tggtccaaga
      181 tgtcgggagc cgcgatcgct gccaaaaagg taagctgacg atagctgacg agattcgatg
      241 cccgacgagc aaatgagcgc cacggctgtt cgcgtaagct cgaaacgacg gagattcgat
      301 agagccagag tcgagtttaa acgattcggg atcttaagga taaatttctg gtaaggagga
      361 cacgtatgga agtgggcaag ttggggaagc cgtatccgtt gctgaatctg gcatatgtgg
      421 gagtataaga cgcgcagcgt cgcatcaggc atttttttct gcgccaatgc acctagaatc
      481 ggttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
      541 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
      601 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
      661 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
      721 cagtctatca actgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttacgc
      781 aaggttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
      841 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
      901 gcggttagct ccttcggtcc tccaatcgtt gtcagaagta agttggccgc agtgttatca
      961 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1021 tctgtgactg gtgaatactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     1081 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     1141 ctcatcattg gaaagcgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     1201 tccagttcga tgtaacccac tcgcgcaccc aactgatctt cagcatcttt tactttcacc
     1261 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     1321 acacggaaat gttgaatact catgcgaaag cctcctgctg atcctgtctc ttgatagatg
     1381 tttgcaaaag ctaggactcg attatacgtt gaccgatcag actatcaata tctaggttta
     1441 aacagatcgg tacgctgccg tgaccaggct tgagtacaat atctgacgat tggacttcct
     1501 gggtcccaag ctggattccg actggctgac tgccaagcaa cgatatggat cgtcgagact
     1561 cgcactgaga ggatcgttct cggtacacca ggcgcgccta gcgttacaca cacaaaaaac
     1621 caacacacat ccatcttcga tggatagcga ttttattatc taagactagg cttacatact
     1681 agcttattcc tttgccctcg gacgagtgct ggggcgtcgg tttccactat cggcgagtac
     1741 ttctacacag ccatcggtcc agacggcggc gcttctgcgg gcgatttgtg tacgcccgac
     1801 agtcccggct ccggatcgga cgattgcgtc gcatcgaccc tgcgcccaag ctgcatcatc
     1861 gaaattgccg tcaaccaagc tctgatagag ttggtcaaga ccaatgcgga gcatatacgc
     1921 ccggagccgg ggcgatcctg caagctccgg atgcctccgc tcgaagtagc gcgtctgctg
     1981 ctccatacaa gccaaccacg gcctccagaa gaagatgttg gcgacctcgt attgggaatc
     2041 cccgaacatc gcctcgctcc agtcaatgac cgctgttatg cggccattgt ccgtcaggac
     2101 attgttggag ccgaaatccg cgtgcacgag gtgccggact tcggggcagt cctcggccca
     2161 aagcatcagc tcatcgagag cctgcgcgac ggacgcactg acggtgtcgt ccatcacagt
     2221 ttgccagtga tacacatggg gatcagcaat cgcgcatatg aaatcacgcc atgtagtgta
     2281 ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc gtctggctaa gatcggcggc
     2341 agcgatggca tccattgcct ccgcgaccgg ctgcagaaca gcgggcagtt cggtttcagg
     2401 caggtcttgc aacgtgacac cctgtgcacg gcgggagatg caataggtca ggctctcgct
     2461 gaactcccca atgtcaagca cttccggaat cgggagcgcg gcggatgcaa agtgccgata
     2521 aacataacga tctttgtaga aaccatcggc gcagctattt acccgcagga catatccacg
     2581 ccctcctaca tcgaagctga aagcacgaga ttcttcgccc tccgagagct gcatcaggtc
     2641 ggagacgctg tcgaactttt cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg
     2701 ctttttaccc atgttgtcta acaaaaaagc caaaaacggc cagaatttag cggacaattt
     2761 acaagtctaa cactgaaaat tacatattga cccaaatgat tacatttcaa aaggtgccta
     2821 aaaaacttca caaaacacac tcgccaaccc cgagcgcata gttcaaaacc ggagcttcag
     2881 ctacttaaga agataggtac ataaaaccga ccaaagaaac tgacgcctca cttatccctc
     2941 ccctcaccag aggtccggcg cctgtcgatt caggagagcc taccctaggc ccgaaccctg
     3001 cgtcctgcga cggagaaaag cctaccgcac acctaccggc tggtggcccc accctgcatt
     3061 ataagccaac agaacgggtg acgtcacgac acgacgaggg cgcgcgctcc caaaggtacg
     3121 ggtgcactgc ccaacggcac cgccataact gccgcccccg caacagacga caaaccgagt
     3181 tctccagtca gtgacaaact tcacgtcagg gtccccagat ggtgcccagc ccatctcacc
     3241 cgaataagag ctttcccgca ttagcgaagg gctcaagacc ttgggttctt gccgcccacc
     3301 atgcccccca ccttgtttca acgacctcac agcccgcctc acaagcgtct tccattcaag
     3361 actcgggaac agccgccatt ttgctgcgct ccccccaacc cccagttcag ggcaaccttg
     3421 ctcgcggacc cagactacag cccttggcgg tctctccaca cgcttccgtc ccaccgagcg
     3481 gcccggcggc cacgaaagcc ccggccagcc cagcagcccg ctactcacca agtgacgatc
     3541 acagcgatcc acaaacaaga accgcgaccc aaatcccggc tgcgacggaa ctagctgtgc
     3601 cacacccggc gcgtccttat ataatcatcg gcgttcaccg ccccacggag atccctccgc
     3661 agaatcgccg agaagggact acttttcctc gcctgttccg ctctctggaa agaaaaccag
     3721 tgccctagag tcacccaagt cccgtcctaa aatgtccttc tgctgatact ggggttctaa
     3781 ggccgagtct tatgagcagc gggccgctgt cctgagcgtc cgggcggaag gatcaggacg
     3841 ctcctgcgcc cttcgtctga cgtggcagcg ctcgccgtga cgaggggggc gcccgcggga
     3901 ggcgccaaaa cccggcgcgg aggccaggcg cgccccgatt tcgatattcc tcggatctcg
     3961 gatagatcgg gattgaggat gtaggttgaa cttctctcgg gtgatcccga tctcggatat
     4021 tcgggattgg agtaatacct ggcgataggc gctgccaagc aatagccaag gcgcaaggcc
     4081 ggccaagaga cttgcgatag ttcgttcgag aggtagcctg caagatggat cggacgaaag
     4141 gtttgaagat tttccgacga gggccttgga gagttcggaa atttaaatgc aactatggat
     4201 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     4261 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4321 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4381 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4441 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4501 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4561 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4621 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4681 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4741 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4801 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4861 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4921 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4981 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5041 ttcctggcct tttgctggcc ttttgctcac atgatttaaa tgaaactcgt aatcggagaa
     5101 cgggtgcaat agcaaggacc agatcgaact tgactgacgc aattctgagc gtagtcgaaa
     5161 cccagacaag tcgatctcgc caagtagctg acttgactgg ttaattaagt ttcgatagcc
     5221 caaggtaacc aatgagatta gggagggaag aaagtttgat gaactacttc tgacccctag
     5281 tggtgtccag aaaagaccat taaaggaatg tcttgatcga aatcgagaga tcgcctgcag
     5341 gatagctgac gaccttaagg ataaatttct ggtaaggagg acacgtatgg aagtgggcaa
     5401 gttggggaag ccgtatccgt tgctgaatct ggcatatgtg ggagtataag acgcgcagcg
     5461 tcgcatcagg catttttttc tgcgccaatg ccggtagata gcaatccggc ccgaggggca
     5521 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctacga taccgataga
     5581 gatgggccaa taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     5641 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     5701 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5761 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtgagctga
     5821 aggtacgctg tatctcagtc agtcaagcta gcagaggcct gcggatcctt attgatggtt
     5881 cgcgtctact ttcggggcct gggcatcatt gagctttttg gcttccgcaa gcaggttggc
     5941 actctgtgat ggatcgtctt ttagggactg gataaatgca tttctttgtt cctcattcag
     6001 attaggaagg tgcaatatct cgtagaaagc attctgctgc tccttgttaa acttgttatc
     6061 aacttttgga gcctgtgcgt cattcagctt cttggcttcg gccaggagat ttgcagactg
     6121 cgagggatcg tctttcaggc tctgaatgaa agcgttgcgt tgctcttcgt tcagatttgg
     6181 taagtgaagg atctcataga aagcgttctg ctgttcttta ttaaacttgt tatcgacggc
     6241 ctcgtcatgt tgtgccaagc caaattcacc ctggaaatag agattctcgg aggcggtcgt
     6301 aggaatgtcg taatctaggg cgccgcttga actgatcttt ttgaaccggt tagccgcaga
     6361 cactgcaata aagttcttct tccatctcct tttccaggga tttatcagcc ccgggattcg
     6421 ccgtcccgac tgaaagtaaa ggttttctgt gagattaaag ttaactacat ctacttgtgc
     6481 tatgtagtta cgagaattca ttctgcacaa gagtagacta tatatcgtaa acggaaaagc
     6541 gaaaacgttt atatagccca tggtggcggc cgctggcggg tgtgtcgagt ggatggtagg
     6601 atcgacaaag atctgatctg acggttcact aaaccagctc tgcttatata gacctcccac
     6661 cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag
     6721 tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga
     6781 aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcagc
     6841 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     6901 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     6961 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     7021 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     7081 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     7141 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctagatctgg
     7201 ctacactcga tcagcagtta gataataaaa
//
