LOCUS       sequence-300594-        6717 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 6717)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6717)
  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-m4252888
FEATURES             Location/Qualifiers
     source          1..6717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    165..236
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(241..287)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    305..353
                     /note="synthetic polyadenylation signal"
     enhancer        434..737
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        739..941
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     892..912
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1010..1019
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1130..1150
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1151..1192
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1261..1382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1272..1291)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1326..1345
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1497..1543
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1573..1709
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1754..1825
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2051..2639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2131..2150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3139..4348
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4185..4204
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4352..5380
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5407..5455
                     /note="synthetic polyadenylation signal"
     CDS             join(5721..5789,5790..6581)
                     /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(5939..5958)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      join(6594..6717,1..13)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc
       61 gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc
      121 agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag
      181 ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt
      241 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt
      301 tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac
      361 tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat
      421 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      481 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      541 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      601 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      661 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      721 tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      781 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      841 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      901 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
      961 tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg ccaccatggg
     1021 ctatataaac gttttcgctt ttccgtttac gatatatagt ctactcttgt gcagaatgaa
     1081 ttctcgtaac tacatagcac aagtagatgt agttaacttt aatctcacag aaaaccttta
     1141 ctttcagtcg gggaaaccta tacccaatcc acttctcggc ttagattcta cctaaggatc
     1201 cgcaggcctc tgctagcttg actgactgag atacagcgta ccttcagctc acagacatga
     1261 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1321 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     1381 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     1441 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
     1501 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
     1561 tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
     1621 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
     1681 tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg
     1741 atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag
     1801 ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat
     1861 taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga
     1921 attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt
     1981 tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2041 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2101 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2161 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2221 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2281 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2341 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2401 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2461 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2521 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2581 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2641 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2701 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2761 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2821 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta
     2881 aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc
     2941 ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct
     3001 attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat
     3061 cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat
     3121 cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc
     3181 tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc
     3241 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     3301 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     3361 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     3421 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     3481 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     3541 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     3601 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     3661 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
     3721 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
     3781 caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc
     3841 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
     3901 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     3961 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
     4021 ccagccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
     4081 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
     4141 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
     4201 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
     4261 gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt
     4321 ctggccgttt ttggcttttt tgttagacaa catgggtaaa aagcctgaac tcaccgcgac
     4381 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     4441 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     4501 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     4561 cgccgcgctc ccgattccgg aagtgcttga cattggggag ttcagcgaga gcctgaccta
     4621 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     4681 cgctgttctg cagccggtcg cggaggcaat ggatgccatc gctgccgccg atcttagcca
     4741 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     4801 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     4861 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     4921 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     4981 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     5041 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     5101 cgagcggagg catccggagc ttgcaggatc gccccggctc cgggcgtata tgctccgcat
     5161 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     5221 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     5281 cgcccgcaga agcgccgccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     5341 aaaccgacgc cccagcactc gtccgagggc aaaggaataa gctagtatgt aagcctagtc
     5401 ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa
     5461 cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     5521 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     5581 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     5641 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     5701 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     5761 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5821 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5881 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     5941 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6001 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     6061 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6121 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6181 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6241 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     6301 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     6361 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6421 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6481 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6541 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     6601 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     6661 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccaga
//
