LOCUS       sequence-300720-        6702 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 6702)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6702)
  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-m4252931
FEATURES             Location/Qualifiers
     source          1..6702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    87..135
                     /note="synthetic polyadenylation signal"
     terminator      153..199
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    204..275
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(427..563)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(576..1367,1368..1436))
                     /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     1199..1218
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1702..1750
                     /note="synthetic polyadenylation signal"
     CDS             complement(1777..2805)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2809..4018)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2953..2972)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4518..5106
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5007..5026
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5332..5403
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5448..5584)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5614..5660)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5775..5896
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5812..5831)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5866..5885
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5965..5991)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(5992..6012)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      6123..6132
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6201..6403)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6230..6250)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(6405..6702,1..6)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactagatc
       61 tggctacact cgatcagcag ttagataata aaatcgctat ccatcgaaga tggatgtgtg
      121 ttggtttttt gtgtgtgtaa cgcaacgatt gatagcataa ccccttgggg cctctaaacg
      181 ggtcttgagg ggttttttga attgagaggt agctgaagct gctgttatga ccactagagg
      241 gaagaagata cctgtggagc taatggtcca agatgtcggg agccgcgatc gctgccaaaa
      301 aggtaagctg acgatagctg acgagattcg atgcccgacg agcaaatgag cgccacggct
      361 gttcgcgtaa gctcgaaacg acggagattc gatagagcca gagtcgagtt taaacgattc
      421 gggatcttaa ggataaattt ctggtaagga ggacacgtat ggaagtgggc aagttgggga
      481 agccgtatcc gttgctgaat ctggcatatg tgggagtata agacgcgcag cgtcgcatca
      541 ggcatttttt tctgcgccaa tgcacctaga atcggttacc aatgcttaat cagtgaggca
      601 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
      661 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
      721 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
      781 agaagtggtc ctgcaacttt atccgcctcc atccagtcta tcaactgttg ccgggaagct
      841 agagtaagta gttcgccagt taatagttta cgcaaggttg ttgccattgc tacaggcatc
      901 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
      961 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccaatc
     1021 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1081 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgaata ctcaaccaag
     1141 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1201 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaagcg ttcttcgggg
     1261 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgcgca
     1321 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1381 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatgcga
     1441 aagcctcctg ctgatcctgt ctcttgatag atgtttgcaa aagctaggac tcgattatac
     1501 gttgaccgat cagactatca atatctaggt ttaaacagat cggtacgctg ccgtgaccag
     1561 gcttgagtac aatatctgac gattggactt cctgggtccc aagctggatt ccgactggct
     1621 gactgccaag caacgatatg gatcgtcgag actcgcactg agaggatcgt tctcggtaca
     1681 ccaggcgcgc ctagcgttac acacacaaaa aaccaacaca catccatctt cgatggatag
     1741 cgattttatt atctaagact aggcttacat actagcttat tcctttgccc tcggacgagt
     1801 gctggggcgt cggtttccac tatcggcgag tacttctaca cagccatcgg tccagacggc
     1861 ggcgcttctg cgggcgattt gtgtacgccc gacagtcccg gctccggatc ggacgattgc
     1921 gtcgcatcga ccctgcgccc aagctgcatc atcgaaattg ccgtcaacca agctctgata
     1981 gagttggtca agaccaatgc ggagcatata cgcccggagc cggggcgatc ctgcaagctc
     2041 cggatgcctc cgctcgaagt agcgcgtctg ctgctccata caagccaacc acggcctcca
     2101 gaagaagatg ttggcgacct cgtattggga atccccgaac atcgcctcgc tccagtcaat
     2161 gaccgctgtt atgcggccat tgtccgtcag gacattgttg gagccgaaat ccgcgtgcac
     2221 gaggtgccgg acttcggggc agtcctcggc ccaaagcatc agctcatcga gagcctgcgc
     2281 gacggacgca ctgacggtgt cgtccatcac agtttgccag tgatacacat ggggatcagc
     2341 aatcgcgcat atgaaatcac gccatgtagt gtattgaccg attccttgcg gtccgaatgg
     2401 gccgaacccg ctcgtctggc taagatcggc ggcagcgatg gcatccattg cctccgcgac
     2461 cggctgcaga acagcgggca gttcggtttc aggcaggtct tgcaacgtga caccctgtgc
     2521 acggcgggag atgcaatagg tcaggctctc gctgaactcc ccaatgtcaa gcacttccgg
     2581 aatcgggagc gcggcggatg caaagtgccg ataaacataa cgatctttgt agaaaccatc
     2641 ggcgcagcta tttacccgca ggacatatcc acgccctcct acatcgaagc tgaaagcacg
     2701 agattcttcg ccctccgaga gctgcatcag gtcggagacg ctgtcgaact tttcgatcag
     2761 aaacttctcg acagacgtcg cggtgagttc aggcttttta cccatgttgt ctaacaaaaa
     2821 agccaaaaac ggccagaatt tagcggacaa tttacaagtc taacactgaa aattacatat
     2881 tgacccaaat gattacattt caaaaggtgc ctaaaaaact tcacaaaaca cactcgccaa
     2941 ccccgagcgc atagttcaaa accggagctt cagctactta agaagatagg tacataaaac
     3001 cgaccaaaga aactgacgcc tcacttatcc ctcccctcac cagaggtccg gcgcctgtcg
     3061 attcaggaga gcctacccta ggcccgaacc ctgcgtcctg cgacggagaa aagcctaccg
     3121 cacacctacc ggctggtggc cccaccctgc attataagcc aacagaacgg gtgacgtcac
     3181 gacacgacga gggcgcgcgc tcccaaaggt acgggtgcac tgcccaacgg caccgccata
     3241 actgccgccc ccgcaacaga cgacaaaccg agttctccag tcagtgacaa acttcacgtc
     3301 agggtcccca gatggtgccc agcccatctc acccgaataa gagctttccc gcattagcga
     3361 agggctcaag accttgggtt cttgccgccc accatgcccc ccaccttgtt tcaacgacct
     3421 cacagcccgc ctcacaagcg tcttccattc aagactcggg aacagccgcc attttgctgc
     3481 gctcccccca acccccagtt cagggcaacc ttgctcgcgg acccagacta cagcccttgg
     3541 cggtctctcc acacgcttcc gtcccaccga gcggcccggc ggccacgaaa gccccggcca
     3601 gcccagcagc ccgctactca ccaagtgacg atcacagcga tccacaaaca agaaccgcga
     3661 cccaaatccc ggctgcgacg gaactagctg tgccacaccc ggcgcgtcct tatataatca
     3721 tcggcgttca ccgccccacg gagatccctc cgcagaatcg ccgagaaggg actacttttc
     3781 ctcgcctgtt ccgctctctg gaaagaaaac cagtgcccta gagtcaccca agtcccgtcc
     3841 taaaatgtcc ttctgctgat actggggttc taaggccgag tcttatgagc agcgggccgc
     3901 tgtcctgagc gtccgggcgg aaggatcagg acgctcctgc gcccttcgtc tgacgtggca
     3961 gcgctcgccg tgacgagggg ggcgcccgcg ggaggcgcca aaacccggcg cggaggccag
     4021 gcgcgccccg atttcgatat tcctcggatc tcggatagat cgggattgag gatgtaggtt
     4081 gaacttctct cgggtgatcc cgatctcgga tattcgggat tggagtaata cctggcgata
     4141 ggcgctgcca agcaatagcc aaggcgcaag gccggccaag agacttgcga tagttcgttc
     4201 gagaggtagc ctgcaagatg gatcggacga aaggtttgaa gattttccga cgagggcctt
     4261 ggagagttcg gaaatttaaa tgcaactatg gatgaacgaa atagacagat cgctgagata
     4321 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4381 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4441 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4501 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4561 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4621 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4681 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4741 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4801 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4861 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4921 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4981 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5041 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5101 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5161 cacatgattt aaatgaaact cgtaatcgga gaacgggtgc aatagcaagg accagatcga
     5221 acttgactga cgcaattctg agcgtagtcg aaacccagac aagtcgatct cgccaagtag
     5281 ctgacttgac tggttaatta agtttcgata gcccaaggta accaatgaga ttagggaggg
     5341 aagaaagttt gatgaactac ttctgacccc tagtggtgtc cagaaaagac cattaaagga
     5401 atgtcttgat cgaaatcgag agatcgcctg caggatagct gacgacctta aggataaatt
     5461 tctggtaagg aggacacgta tggaagtggg caagttgggg aagccgtatc cgttgctgaa
     5521 tctggcatat gtgggagtat aagacgcgca gcgtcgcatc aggcattttt ttctgcgcca
     5581 atgccggtag atagcaatcc ggcccgaggg gcacaaaaaa cccctcaaga cccgtttaga
     5641 ggccccaagg ggttatgcta cgataccgat agagatgggc caataccaca tttgtagagg
     5701 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     5761 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5821 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5881 tcatcaatgt atcttatcat gtctgtgagc tgaaggtacg ctgtatctca gtcagtcaag
     5941 ctagcagagg cctgcggatc cttacgcata atctggtacg tcataggggt acgactgaaa
     6001 gtaaaggttt tctgtgagat taaagttaac tacatctact tgtgctatgt agttacgaga
     6061 attcattctg cacaagagta gactatatat cgtaaacgga aaagcgaaaa cgtttatata
     6121 gcccatggtg gcggccgctg gcgggtgtgt cgagtggatg gtaggatcga caaagatctg
     6181 atctgacggt tcactaaacc agctctgctt atatagacct cccaccgtac acgcctaccg
     6241 cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg
     6301 tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca
     6361 aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcagcatggt aatagcgatg
     6421 actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa
     6481 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac
     6541 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg
     6601 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg
     6661 tcgttgggcg gtcagccagg cgggccattt accgtaagtt at
//
