LOCUS       sequence-300763-        6744 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 6744)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6744)
  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-m4252879
FEATURES             Location/Qualifiers
     source          1..6744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    162..233
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(459..1047)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(539..558)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1547..2756
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2593..2612
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             2760..3788
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    3815..3863
                     /note="synthetic polyadenylation signal"
     CDS             join(4129..4197,4198..4989)
                     /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(4347..4366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5002..5138
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5290..5361
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5366..5412)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5430..5478
                     /note="synthetic polyadenylation signal"
     enhancer        5559..5862
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5864..6066
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6017..6037
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6135..6144
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6255..6275
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             6279..6344
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    6413..6534
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6424..6443)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6478..6497
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      6649..6695
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      join(6725..6744,1..117)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
       61 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt
      121 cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt
      181 tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat
      241 tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg
      301 acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc
      361 tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa
      421 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
      481 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      541 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      601 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
      661 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      721 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      781 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      841 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
      901 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
      961 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1021 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1081 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     1141 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1201 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     1261 ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa
     1321 aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa
     1381 gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac
     1441 tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat
     1501 cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg
     1561 gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa
     1621 gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat
     1681 aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg
     1741 actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc
     1801 tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg
     1861 cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg
     1921 tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt
     1981 ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct
     2041 gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc
     2101 tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag
     2161 gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa
     2221 gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca
     2281 ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg
     2341 ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc
     2401 tgttggctta taatgcaggg tggggccacc agccggtagg tgtgcggtag gcttttctcc
     2461 gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc
     2521 tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc
     2581 ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt
     2641 gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt
     2701 gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca
     2761 tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg
     2821 acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg
     2881 atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag
     2941 atcgttatgt ttatcggcac tttgcatccg ccgcgctccc gattccggaa gtgcttgaca
     3001 ttggggagtt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt
     3061 tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggcaatgg
     3121 atgccatcgc tgccgccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag
     3181 gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt
     3241 atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg
     3301 agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg
     3361 gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg
     3421 cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg
     3481 cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc
     3541 cccggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg
     3601 acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg
     3661 gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgccgccgtc tggaccgatg
     3721 gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa
     3781 aggaataagc tagtatgtaa gcctagtctt agataataaa atcgctatcc atcgaagatg
     3841 gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc
     3901 ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca
     3961 gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt
     4021 accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta
     4081 gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa
     4141 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4201 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac
     4261 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt
     4321 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4381 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca
     4441 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4501 ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag
     4561 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4621 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4681 gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag
     4741 ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4801 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4861 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4921 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4981 cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc
     5041 gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac
     5101 ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg tttaaactcg
     5161 actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt
     5221 tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc
     5281 ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta gtggtcataa
     5341 cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta gaggccccaa
     5401 ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca tccatcttcg
     5461 atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta gtaatcaatt
     5521 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     5581 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     5641 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     5701 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     5761 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     5821 acttggcagt acatctacgt attagtcatc gctattacca tgctgatgcg gttttggcag
     5881 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     5941 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     6001 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     6061 agagctggtt tagtgaaccg tcagatcaga tctttgtcga tcctaccatc cactcgacac
     6121 acccgccagc ggccgccacc atggggtaca taaatgtttt cgcctttcca ttcacgatct
     6181 attcactcct tttatgtagg atgaactctc gcaattacat tgctcaggta gacgtcgtga
     6241 actttaacct aactgaaaac ctttactttc agtcgatgga ttataaggac cacgatggag
     6301 actacaaaga tcatgacatt gattacaagg acgacgatga caaataagga tccgcaggcc
     6361 tctgctagct tgactgactg agatacagcg taccttcagc tcacagacat gataagatac
     6421 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     6481 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     6541 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     6601 aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta gcataacccc
     6661 ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg attgctatct
     6721 accggcattg gcgcagaaaa aaat
//
