LOCUS       sequence-307358-        7065 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 7065)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7065)
  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-m4252913
FEATURES             Location/Qualifiers
     source          1..7065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..31
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(135..152)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(501..521)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      632..641
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(710..912)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(739..759)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        914..1217
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    1298..1346
                     /note="synthetic polyadenylation signal"
     terminator      1364..1410
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1415..1486
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1638..1774)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(1787..2578,2579..2647))
                     /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     2410..2429
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    2913..2961
                     /note="synthetic polyadenylation signal"
     CDS             complement(2988..4016)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(4020..5229)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(4164..4183)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      5729..6317
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6218..6237
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6543..6614
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6659..6795)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(6825..6871)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    join(6986..7065,1..42)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7023..7042)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtgagctgaa
       61 ggtacgctgt atctcagtca gtcaagctag cagaggcctg cggatcctta gccgcgtggc
      121 acgagcccgc tgctatggtg atggtgatgg tgcatgtgcc ccgagcctga acctgccagg
      181 ttagcgtcca agaactcttt tagctggccc ttagacaggg ctcccacttt tgtcgcggca
      241 acttccccat tcttaaagag caggagggta ggtattcccc tgatgccgta tttaggtgcc
      301 gtccctgggt tctgatctat attaagctta gcgacagtta attttccttg atactcgtcg
      361 gcgatctcgt ccagaatggg tgcgatcatc ttgcacgggc cacaccattc agcccagaaa
      421 tctaccagta tcgcaccatc ggccttcaac acgtctgtat caaaggagtc atcggttaga
      481 tgaatgattt tgtcactcat cgactgaaag taaaggtttt ctgtgagatt aaagttaact
      541 acatctactt gtgctatgta gttacgagaa ttcattctgc acaagagtag actatatatc
      601 gtaaacggaa aagcgaaaac gtttatatag cccatggtgg cggccgctgg cgggtgtgtc
      661 gagtggatgg taggatcgac aaagatctga tctgacggtt cactaaacca gctctgctta
      721 tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac
      781 gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg
      841 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc
      901 aaaaccgcat cagcatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
      961 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     1021 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     1081 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     1141 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     1201 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     1261 attactagat ctggctacac tcgatcagca gttagataat aaaatcgcta tccatcgaag
     1321 atggatgtgt gttggttttt tgtgtgtgta acgcaacgat tgatagcata accccttggg
     1381 gcctctaaac gggtcttgag gggttttttg aattgagagg tagctgaagc tgctgttatg
     1441 accactagag ggaagaagat acctgtggag ctaatggtcc aagatgtcgg gagccgcgat
     1501 cgctgccaaa aaggtaagct gacgatagct gacgagattc gatgcccgac gagcaaatga
     1561 gcgccacggc tgttcgcgta agctcgaaac gacggagatt cgatagagcc agagtcgagt
     1621 ttaaacgatt cgggatctta aggataaatt tctggtaagg aggacacgta tggaagtggg
     1681 caagttgggg aagccgtatc cgttgctgaa tctggcatat gtgggagtat aagacgcgca
     1741 gcgtcgcatc aggcattttt ttctgcgcca atgcacctag aatcggttac caatgcttaa
     1801 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     1861 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     1921 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     1981 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct atcaactgtt
     2041 gccgggaagc tagagtaagt agttcgccag ttaatagttt acgcaaggtt gttgccattg
     2101 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     2161 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     2221 gtcctccaat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     2281 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgaat
     2341 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     2401 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaagc
     2461 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     2521 ccactcgcgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     2581 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     2641 tactcatgcg aaagcctcct gctgatcctg tctcttgata gatgtttgca aaagctagga
     2701 ctcgattata cgttgaccga tcagactatc aatatctagg tttaaacaga tcggtacgct
     2761 gccgtgacca ggcttgagta caatatctga cgattggact tcctgggtcc caagctggat
     2821 tccgactggc tgactgccaa gcaacgatat ggatcgtcga gactcgcact gagaggatcg
     2881 ttctcggtac accaggcgcg cctagcgtta cacacacaaa aaaccaacac acatccatct
     2941 tcgatggata gcgattttat tatctaagac taggcttaca tactagctta ttcctttgcc
     3001 ctcggacgag tgctggggcg tcggtttcca ctatcggcga gtacttctac acagccatcg
     3061 gtccagacgg cggcgcttct gcgggcgatt tgtgtacgcc cgacagtccc ggctccggat
     3121 cggacgattg cgtcgcatcg accctgcgcc caagctgcat catcgaaatt gccgtcaacc
     3181 aagctctgat agagttggtc aagaccaatg cggagcatat acgcccggag ccggggcgat
     3241 cctgcaagct ccggatgcct ccgctcgaag tagcgcgtct gctgctccat acaagccaac
     3301 cacggcctcc agaagaagat gttggcgacc tcgtattggg aatccccgaa catcgcctcg
     3361 ctccagtcaa tgaccgctgt tatgcggcca ttgtccgtca ggacattgtt ggagccgaaa
     3421 tccgcgtgca cgaggtgccg gacttcgggg cagtcctcgg cccaaagcat cagctcatcg
     3481 agagcctgcg cgacggacgc actgacggtg tcgtccatca cagtttgcca gtgatacaca
     3541 tggggatcag caatcgcgca tatgaaatca cgccatgtag tgtattgacc gattccttgc
     3601 ggtccgaatg ggccgaaccc gctcgtctgg ctaagatcgg cggcagcgat ggcatccatt
     3661 gcctccgcga ccggctgcag aacagcgggc agttcggttt caggcaggtc ttgcaacgtg
     3721 acaccctgtg cacggcggga gatgcaatag gtcaggctct cgctgaactc cccaatgtca
     3781 agcacttccg gaatcgggag cgcggcggat gcaaagtgcc gataaacata acgatctttg
     3841 tagaaaccat cggcgcagct atttacccgc aggacatatc cacgccctcc tacatcgaag
     3901 ctgaaagcac gagattcttc gccctccgag agctgcatca ggtcggagac gctgtcgaac
     3961 ttttcgatca gaaacttctc gacagacgtc gcggtgagtt caggcttttt acccatgttg
     4021 tctaacaaaa aagccaaaaa cggccagaat ttagcggaca atttacaagt ctaacactga
     4081 aaattacata ttgacccaaa tgattacatt tcaaaaggtg cctaaaaaac ttcacaaaac
     4141 acactcgcca accccgagcg catagttcaa aaccggagct tcagctactt aagaagatag
     4201 gtacataaaa ccgaccaaag aaactgacgc ctcacttatc cctcccctca ccagaggtcc
     4261 ggcgcctgtc gattcaggag agcctaccct aggcccgaac cctgcgtcct gcgacggaga
     4321 aaagcctacc gcacacctac cggctggtgg ccccaccctg cattataagc caacagaacg
     4381 ggtgacgtca cgacacgacg agggcgcgcg ctcccaaagg tacgggtgca ctgcccaacg
     4441 gcaccgccat aactgccgcc cccgcaacag acgacaaacc gagttctcca gtcagtgaca
     4501 aacttcacgt cagggtcccc agatggtgcc cagcccatct cacccgaata agagctttcc
     4561 cgcattagcg aagggctcaa gaccttgggt tcttgccgcc caccatgccc cccaccttgt
     4621 ttcaacgacc tcacagcccg cctcacaagc gtcttccatt caagactcgg gaacagccgc
     4681 cattttgctg cgctcccccc aacccccagt tcagggcaac cttgctcgcg gacccagact
     4741 acagcccttg gcggtctctc cacacgcttc cgtcccaccg agcggcccgg cggccacgaa
     4801 agccccggcc agcccagcag cccgctactc accaagtgac gatcacagcg atccacaaac
     4861 aagaaccgcg acccaaatcc cggctgcgac ggaactagct gtgccacacc cggcgcgtcc
     4921 ttatataatc atcggcgttc accgccccac ggagatccct ccgcagaatc gccgagaagg
     4981 gactactttt cctcgcctgt tccgctctct ggaaagaaaa ccagtgccct agagtcaccc
     5041 aagtcccgtc ctaaaatgtc cttctgctga tactggggtt ctaaggccga gtcttatgag
     5101 cagcgggccg ctgtcctgag cgtccgggcg gaaggatcag gacgctcctg cgcccttcgt
     5161 ctgacgtggc agcgctcgcc gtgacgaggg gggcgcccgc gggaggcgcc aaaacccggc
     5221 gcggaggcca ggcgcgcccc gatttcgata ttcctcggat ctcggataga tcgggattga
     5281 ggatgtaggt tgaacttctc tcgggtgatc ccgatctcgg atattcggga ttggagtaat
     5341 acctggcgat aggcgctgcc aagcaatagc caaggcgcaa ggccggccaa gagacttgcg
     5401 atagttcgtt cgagaggtag cctgcaagat ggatcggacg aaaggtttga agattttccg
     5461 acgagggcct tggagagttc ggaaatttaa atgcaactat ggatgaacga aatagacaga
     5521 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5581 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5641 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5701 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5761 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5821 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5881 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5941 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6001 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6061 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6121 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6181 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6241 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6301 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6361 ggccttttgc tcacatgatt taaatgaaac tcgtaatcgg agaacgggtg caatagcaag
     6421 gaccagatcg aacttgactg acgcaattct gagcgtagtc gaaacccaga caagtcgatc
     6481 tcgccaagta gctgacttga ctggttaatt aagtttcgat agcccaaggt aaccaatgag
     6541 attagggagg gaagaaagtt tgatgaacta cttctgaccc ctagtggtgt ccagaaaaga
     6601 ccattaaagg aatgtcttga tcgaaatcga gagatcgcct gcaggatagc tgacgacctt
     6661 aaggataaat ttctggtaag gaggacacgt atggaagtgg gcaagttggg gaagccgtat
     6721 ccgttgctga atctggcata tgtgggagta taagacgcgc agcgtcgcat caggcatttt
     6781 tttctgcgcc aatgccggta gatagcaatc cggcccgagg ggcacaaaaa acccctcaag
     6841 acccgtttag aggccccaag gggttatgct acgataccga tagagatggg ccaataccac
     6901 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     6961 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     7021 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcact
//
