LOCUS       sequence-300721-        6753 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 6753)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6753)
  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-m4252956
FEATURES             Location/Qualifiers
     source          1..6753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      234..822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     723..742
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1048..1119
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1164..1300)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(1330..1376)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    1491..1612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1528..1547)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1582..1601
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1681..1758)
                     /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(1759..1779)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      1890..1899
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1968..2170)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1997..2017)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2172..2475
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    2556..2604
                     /note="synthetic polyadenylation signal"
     terminator      2622..2668
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2673..2744
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2896..3032)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(3045..3836,3837..3905))
                     /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     3668..3687
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    4171..4219
                     /note="synthetic polyadenylation signal"
     CDS             complement(4246..5274)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(5278..6487)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5422..5441)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
       61 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
      121 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
      181 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
      241 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
      301 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
      361 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
      421 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
      481 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
      541 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
      601 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
      661 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
      721 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
      781 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      841 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgatttaaat gaaactcgta
      901 atcggagaac gggtgcaata gcaaggacca gatcgaactt gactgacgca attctgagcg
      961 tagtcgaaac ccagacaagt cgatctcgcc aagtagctga cttgactggt taattaagtt
     1021 tcgatagccc aaggtaacca atgagattag ggagggaaga aagtttgatg aactacttct
     1081 gacccctagt ggtgtccaga aaagaccatt aaaggaatgt cttgatcgaa atcgagagat
     1141 cgcctgcagg atagctgacg accttaagga taaatttctg gtaaggagga cacgtatgga
     1201 agtgggcaag ttggggaagc cgtatccgtt gctgaatctg gcatatgtgg gagtataaga
     1261 cgcgcagcgt cgcatcaggc atttttttct gcgccaatgc cggtagatag caatccggcc
     1321 cgaggggcac aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctacgat
     1381 accgatagag atgggccaat accacatttg tagaggtttt acttgcttta aaaaacctcc
     1441 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     1501 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1561 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     1621 gtgagctgaa ggtacgctgt atctcagtca gtcaagctag cagaggcctg cggatcctta
     1681 gagggcacca gacgaggata tctttttgaa tctgttggct gcactcaccg cgataaaatt
     1741 ctttttccat cgcctcttcg actgaaagta aaggttttca gttaggttaa agttcacgac
     1801 gtctacctga gcaatgtaat tgcgagagtt catcctacat aaaaggagtg aatagatcgt
     1861 gaatggaaag gcgaaaacat ttatgtaccc catggtggcg gccgctggcg ggtgtgtcga
     1921 gtggatggta ggatcgacaa agatctgatc tgacggttca ctaaaccagc tctgcttata
     1981 tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga
     2041 cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg
     2101 tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa
     2161 aaccgcatca gcatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag
     2221 tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca
     2281 atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt
     2341 aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac
     2401 gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc
     2461 gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat
     2521 tactagatct ggctacactc gatcagcagt tagataataa aatcgctatc catcgaagat
     2581 ggatgtgtgt tggttttttg tgtgtgtaac gcaacgattg atagcataac cccttggggc
     2641 ctctaaacgg gtcttgaggg gttttttgaa ttgagaggta gctgaagctg ctgttatgac
     2701 cactagaggg aagaagatac ctgtggagct aatggtccaa gatgtcggga gccgcgatcg
     2761 ctgccaaaaa ggtaagctga cgatagctga cgagattcga tgcccgacga gcaaatgagc
     2821 gccacggctg ttcgcgtaag ctcgaaacga cggagattcg atagagccag agtcgagttt
     2881 aaacgattcg ggatcttaag gataaatttc tggtaaggag gacacgtatg gaagtgggca
     2941 agttggggaa gccgtatccg ttgctgaatc tggcatatgt gggagtataa gacgcgcagc
     3001 gtcgcatcag gcattttttt ctgcgccaat gcacctagaa tcggttacca atgcttaatc
     3061 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3121 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3181 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3241 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat caactgttgc
     3301 cgggaagcta gagtaagtag ttcgccagtt aatagtttac gcaaggttgt tgccattgct
     3361 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3421 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3481 cctccaatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3541 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgaatac
     3601 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3661 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaagcgt
     3721 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3781 actcgcgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3841 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3901 ctcatgcgaa agcctcctgc tgatcctgtc tcttgataga tgtttgcaaa agctaggact
     3961 cgattatacg ttgaccgatc agactatcaa tatctaggtt taaacagatc ggtacgctgc
     4021 cgtgaccagg cttgagtaca atatctgacg attggacttc ctgggtccca agctggattc
     4081 cgactggctg actgccaagc aacgatatgg atcgtcgaga ctcgcactga gaggatcgtt
     4141 ctcggtacac caggcgcgcc tagcgttaca cacacaaaaa accaacacac atccatcttc
     4201 gatggatagc gattttatta tctaagacta ggcttacata ctagcttatt cctttgccct
     4261 cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac agccatcggt
     4321 ccagacggcg gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg ctccggatcg
     4381 gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa
     4441 gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc ggggcgatcc
     4501 tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac aagccaacca
     4561 cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca tcgcctcgct
     4621 ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg agccgaaatc
     4681 cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca gctcatcgag
     4741 agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt gatacacatg
     4801 gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg
     4861 tccgaatggg ccgaacccgc tcgtctggct aagatcggcg gcagcgatgg catccattgc
     4921 ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac
     4981 accctgtgca cggcgggaga tgcaataggt caggctctcg ctgaactccc caatgtcaag
     5041 cacttccgga atcgggagcg cggcggatgc aaagtgccga taaacataac gatctttgta
     5101 gaaaccatcg gcgcagctat ttacccgcag gacatatcca cgccctccta catcgaagct
     5161 gaaagcacga gattcttcgc cctccgagag ctgcatcagg tcggagacgc tgtcgaactt
     5221 ttcgatcaga aacttctcga cagacgtcgc ggtgagttca ggctttttac ccatgttgtc
     5281 taacaaaaaa gccaaaaacg gccagaattt agcggacaat ttacaagtct aacactgaaa
     5341 attacatatt gacccaaatg attacatttc aaaaggtgcc taaaaaactt cacaaaacac
     5401 actcgccaac cccgagcgca tagttcaaaa ccggagcttc agctacttaa gaagataggt
     5461 acataaaacc gaccaaagaa actgacgcct cacttatccc tcccctcacc agaggtccgg
     5521 cgcctgtcga ttcaggagag cctaccctag gcccgaaccc tgcgtcctgc gacggagaaa
     5581 agcctaccgc acacctaccg gctggtggcc ccaccctgca ttataagcca acagaacggg
     5641 tgacgtcacg acacgacgag ggcgcgcgct cccaaaggta cgggtgcact gcccaacggc
     5701 accgccataa ctgccgcccc cgcaacagac gacaaaccga gttctccagt cagtgacaaa
     5761 cttcacgtca gggtccccag atggtgccca gcccatctca cccgaataag agctttcccg
     5821 cattagcgaa gggctcaaga ccttgggttc ttgccgccca ccatgccccc caccttgttt
     5881 caacgacctc acagcccgcc tcacaagcgt cttccattca agactcggga acagccgcca
     5941 ttttgctgcg ctccccccaa cccccagttc agggcaacct tgctcgcgga cccagactac
     6001 agcccttggc ggtctctcca cacgcttccg tcccaccgag cggcccggcg gccacgaaag
     6061 ccccggccag cccagcagcc cgctactcac caagtgacga tcacagcgat ccacaaacaa
     6121 gaaccgcgac ccaaatcccg gctgcgacgg aactagctgt gccacacccg gcgcgtcctt
     6181 atataatcat cggcgttcac cgccccacgg agatccctcc gcagaatcgc cgagaaggga
     6241 ctacttttcc tcgcctgttc cgctctctgg aaagaaaacc agtgccctag agtcacccaa
     6301 gtcccgtcct aaaatgtcct tctgctgata ctggggttct aaggccgagt cttatgagca
     6361 gcgggccgct gtcctgagcg tccgggcgga aggatcagga cgctcctgcg cccttcgtct
     6421 gacgtggcag cgctcgccgt gacgaggggg gcgcccgcgg gaggcgccaa aacccggcgc
     6481 ggaggccagg cgcgccccga tttcgatatt cctcggatct cggatagatc gggattgagg
     6541 atgtaggttg aacttctctc gggtgatccc gatctcggat attcgggatt ggagtaatac
     6601 ctggcgatag gcgctgccaa gcaatagcca aggcgcaagg ccggccaaga gacttgcgat
     6661 agttcgttcg agaggtagcc tgcaagatgg atcggacgaa aggtttgaag attttccgac
     6721 gagggccttg gagagttcgg aaatttaaat gca
//
