LOCUS       sequence-307387-        4392 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 4392)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4392)
  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-m4133365
FEATURES             Location/Qualifiers
     source          1..4392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      256..392
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    544..615
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(620..666)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    684..732
                     /note="synthetic polyadenylation signal"
     enhancer        813..1116
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1118..1320
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1271..1291
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1389..1398
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1509..1529
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1533..1598
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1651..1772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1662..1681)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1716..1735
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1887..1933
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1963..2099
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2144..2215
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2441..3029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2521..2540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3775..3843,3844..4392,1..243)
                     /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(3993..4012)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
       61 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
      121 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
      181 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
      241 taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta
      301 tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat
      361 acgtgtcctc cttaccagaa atttatcctt aagatcccga atcgtttaaa ctcgactctg
      421 gctctatcga atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg
      481 tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc
      541 cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag
      601 cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt
      661 atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat
      721 agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg
      781 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      841 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      901 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      961 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     1021 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     1081 cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc
     1141 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     1201 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     1261 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     1321 ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc
     1381 cagcggccgc caccatgggg tacataaatg ttttcgcctt tccattcacg atctattcac
     1441 tccttttatg taggatgaac tctcgcaatt acattgctca ggtagacgtc gtgaacttta
     1501 acctaactga aaacctttac tttcagtcga tggattataa ggaccacgat ggagactaca
     1561 aagatcatga cattgattac aaggacgacg atgacaaata agctagcttg actgactgag
     1621 atacagcgta ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca
     1681 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     1741 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     1801 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     1861 gtattggccc atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt
     1921 gaggggtttt ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa
     1981 atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata
     2041 cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag
     2101 gtcgtcagct atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct
     2161 tttctggaca ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc
     2221 attggttacc ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat
     2281 cgacttgtct gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt
     2341 gctattgcac ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc
     2401 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2461 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2521 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2581 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2641 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2701 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2761 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2821 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2881 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2941 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3001 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3061 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3121 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3181 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3241 gtctatttcg ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg
     3301 aaaatcttca aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc
     3361 aagtctcttg gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt
     3421 actccaatcc cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca
     3481 atcccgatct atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg tgtaccgaga
     3541 acgatcctct cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg
     3601 aatccagctt gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg
     3661 cagcgtaccg atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga
     3721 gtcctagctt ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt
     3781 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3841 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg
     3901 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3961 cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4021 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4081 tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4141 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga
     4201 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4261 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4321 gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg
     4381 caacagttga ta
//
