LOCUS       sequence-300144-        4350 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 4350)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4350)
  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-m4133705
FEATURES             Location/Qualifiers
     source          1..4350
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             119..139
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             140..166
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    219..340
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(230..249)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     284..303
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      455..501
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      531..667
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    712..783
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1009..1597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1089..1108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2343..2411,2412..3203)
                     /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(2561..2580)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3216..3352
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3504..3575
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3580..3626)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3644..3692
                     /note="synthetic polyadenylation signal"
     enhancer        3773..4076
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4078..4280
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4231..4251
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4349..4350,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatgggc tatataaacg ttttcgcttt tccgtttacg atatatagtc tactcttgtg
       61 cagaatgaat tctcgtaact acatagcaca agtagatgta gttaacttta atctcacaga
      121 aaacctttac tttcagtcgt acccctatga cgtaccagat tatgcgtaag ctagcttgac
      181 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
      241 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      301 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      361 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
      421 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
      481 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
      541 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
      601 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
      661 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
      721 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
      781 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
      841 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
      901 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
      961 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1021 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1081 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1141 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1201 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1261 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1321 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1381 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1441 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1501 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1561 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1621 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1681 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1741 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1801 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     1861 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     1921 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     1981 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     2041 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg
     2101 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     2161 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     2221 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     2281 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     2341 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2401 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2461 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2521 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2581 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2641 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2701 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2761 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2821 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2881 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     2941 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     3001 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3061 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3121 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3181 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     3241 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     3301 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga
     3361 atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag
     3421 ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt
     3481 ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc
     3541 tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg
     3601 tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca
     3661 cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga
     3721 tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     3781 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     3841 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     3901 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     3961 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     4021 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga
     4081 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     4141 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     4201 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     4261 aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac
     4321 catccactcg acacacccgc cagcggccgc
//
