LOCUS       sequence-300429-        6654 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 6654)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6654)
  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-m4252990
FEATURES             Location/Qualifiers
     source          1..6654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1165..1753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1654..1673
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1979..2050
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2095..2231)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(2261..2307)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    2422..2543
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2459..2478)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2513..2532
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     regulatory      2722..2731
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2800..3002)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2829..2849)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3004..3307
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    3388..3436
                     /note="synthetic polyadenylation signal"
     terminator      3454..3500
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    3505..3576
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3728..3864)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(3877..4668,4669..4737))
                     /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     4500..4519
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5003..5051
                     /note="synthetic polyadenylation signal"
     CDS             complement(5078..6106)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(join(6110..6654,1..665))
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(6254..6273)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 ttagcgaagg gctcaagacc ttgggttctt gccgcccacc atgcccccca ccttgtttca
       61 acgacctcac agcccgcctc acaagcgtct tccattcaag actcgggaac agccgccatt
      121 ttgctgcgct ccccccaacc cccagttcag ggcaaccttg ctcgcggacc cagactacag
      181 cccttggcgg tctctccaca cgcttccgtc ccaccgagcg gcccggcggc cacgaaagcc
      241 ccggccagcc cagcagcccg ctactcacca agtgacgatc acagcgatcc acaaacaaga
      301 accgcgaccc aaatcccggc tgcgacggaa ctagctgtgc cacacccggc gcgtccttat
      361 ataatcatcg gcgttcaccg ccccacggag atccctccgc agaatcgccg agaagggact
      421 acttttcctc gcctgttccg ctctctggaa agaaaaccag tgccctagag tcacccaagt
      481 cccgtcctaa aatgtccttc tgctgatact ggggttctaa ggccgagtct tatgagcagc
      541 gggccgctgt cctgagcgtc cgggcggaag gatcaggacg ctcctgcgcc cttcgtctga
      601 cgtggcagcg ctcgccgtga cgaggggggc gcccgcggga ggcgccaaaa cccggcgcgg
      661 aggccaggcg cgccccgatt tcgatattcc tcggatctcg gatagatcgg gattgaggat
      721 gtaggttgaa cttctctcgg gtgatcccga tctcggatat tcgggattgg agtaatacct
      781 ggcgataggc gctgccaagc aatagccaag gcgcaaggcc ggccaagaga cttgcgatag
      841 ttcgttcgag aggtagcctg caagatggat cggacgaaag gtttgaagat tttccgacga
      901 gggccttgga gagttcggaa atttaaatgc aactatggat gaacgaaata gacagatcgc
      961 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     1021 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1081 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1141 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1201 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1261 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1321 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1381 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1441 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1501 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1561 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1621 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1681 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1741 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1801 ttttgctcac atgatttaaa tgaaactcgt aatcggagaa cgggtgcaat agcaaggacc
     1861 agatcgaact tgactgacgc aattctgagc gtagtcgaaa cccagacaag tcgatctcgc
     1921 caagtagctg acttgactgg ttaattaagt ttcgatagcc caaggtaacc aatgagatta
     1981 gggagggaag aaagtttgat gaactacttc tgacccctag tggtgtccag aaaagaccat
     2041 taaaggaatg tcttgatcga aatcgagaga tcgcctgcag gatagctgac gaccttaagg
     2101 ataaatttct ggtaaggagg acacgtatgg aagtgggcaa gttggggaag ccgtatccgt
     2161 tgctgaatct ggcatatgtg ggagtataag acgcgcagcg tcgcatcagg catttttttc
     2221 tgcgccaatg ccggtagata gcaatccggc ccgaggggca caaaaaaccc ctcaagaccc
     2281 gtttagaggc cccaaggggt tatgctacga taccgataga gatgggccaa taccacattt
     2341 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     2401 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     2461 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2521 tccaaactca tcaatgtatc ttatcatgtc tgtgagctga aggtacgctg tatctcagtc
     2581 agtcaagcta gcagaggcct gcggatcctt aagttaggtt aaagttcacg acgtctacct
     2641 gagcaatgta attgcgagag ttcatcctac ataaaaggag tgaatagatc gtgaatggaa
     2701 aggcgaaaac atttatgtac cccatggtgg cggccgctgg cgggtgtgtc gagtggatgg
     2761 taggatcgac aaagatctga tctgacggtt cactaaacca gctctgctta tatagacctc
     2821 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg
     2881 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact
     2941 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat
     3001 cagcatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
     3061 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
     3121 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
     3181 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
     3241 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
     3301 tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg attactagat
     3361 ctggctacac tcgatcagca gttagataat aaaatcgcta tccatcgaag atggatgtgt
     3421 gttggttttt tgtgtgtgta acgcaacgat tgatagcata accccttggg gcctctaaac
     3481 gggtcttgag gggttttttg aattgagagg tagctgaagc tgctgttatg accactagag
     3541 ggaagaagat acctgtggag ctaatggtcc aagatgtcgg gagccgcgat cgctgccaaa
     3601 aaggtaagct gacgatagct gacgagattc gatgcccgac gagcaaatga gcgccacggc
     3661 tgttcgcgta agctcgaaac gacggagatt cgatagagcc agagtcgagt ttaaacgatt
     3721 cgggatctta aggataaatt tctggtaagg aggacacgta tggaagtggg caagttgggg
     3781 aagccgtatc cgttgctgaa tctggcatat gtgggagtat aagacgcgca gcgtcgcatc
     3841 aggcattttt ttctgcgcca atgcacctag aatcggttac caatgcttaa tcagtgaggc
     3901 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3961 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4021 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4081 cagaagtggt cctgcaactt tatccgcctc catccagtct atcaactgtt gccgggaagc
     4141 tagagtaagt agttcgccag ttaatagttt acgcaaggtt gttgccattg ctacaggcat
     4201 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4261 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccaat
     4321 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4381 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgaat actcaaccaa
     4441 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4501 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaagc gttcttcggg
     4561 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgcgc
     4621 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4681 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatgcg
     4741 aaagcctcct gctgatcctg tctcttgata gatgtttgca aaagctagga ctcgattata
     4801 cgttgaccga tcagactatc aatatctagg tttaaacaga tcggtacgct gccgtgacca
     4861 ggcttgagta caatatctga cgattggact tcctgggtcc caagctggat tccgactggc
     4921 tgactgccaa gcaacgatat ggatcgtcga gactcgcact gagaggatcg ttctcggtac
     4981 accaggcgcg cctagcgtta cacacacaaa aaaccaacac acatccatct tcgatggata
     5041 gcgattttat tatctaagac taggcttaca tactagctta ttcctttgcc ctcggacgag
     5101 tgctggggcg tcggtttcca ctatcggcga gtacttctac acagccatcg gtccagacgg
     5161 cggcgcttct gcgggcgatt tgtgtacgcc cgacagtccc ggctccggat cggacgattg
     5221 cgtcgcatcg accctgcgcc caagctgcat catcgaaatt gccgtcaacc aagctctgat
     5281 agagttggtc aagaccaatg cggagcatat acgcccggag ccggggcgat cctgcaagct
     5341 ccggatgcct ccgctcgaag tagcgcgtct gctgctccat acaagccaac cacggcctcc
     5401 agaagaagat gttggcgacc tcgtattggg aatccccgaa catcgcctcg ctccagtcaa
     5461 tgaccgctgt tatgcggcca ttgtccgtca ggacattgtt ggagccgaaa tccgcgtgca
     5521 cgaggtgccg gacttcgggg cagtcctcgg cccaaagcat cagctcatcg agagcctgcg
     5581 cgacggacgc actgacggtg tcgtccatca cagtttgcca gtgatacaca tggggatcag
     5641 caatcgcgca tatgaaatca cgccatgtag tgtattgacc gattccttgc ggtccgaatg
     5701 ggccgaaccc gctcgtctgg ctaagatcgg cggcagcgat ggcatccatt gcctccgcga
     5761 ccggctgcag aacagcgggc agttcggttt caggcaggtc ttgcaacgtg acaccctgtg
     5821 cacggcggga gatgcaatag gtcaggctct cgctgaactc cccaatgtca agcacttccg
     5881 gaatcgggag cgcggcggat gcaaagtgcc gataaacata acgatctttg tagaaaccat
     5941 cggcgcagct atttacccgc aggacatatc cacgccctcc tacatcgaag ctgaaagcac
     6001 gagattcttc gccctccgag agctgcatca ggtcggagac gctgtcgaac ttttcgatca
     6061 gaaacttctc gacagacgtc gcggtgagtt caggcttttt acccatgttg tctaacaaaa
     6121 aagccaaaaa cggccagaat ttagcggaca atttacaagt ctaacactga aaattacata
     6181 ttgacccaaa tgattacatt tcaaaaggtg cctaaaaaac ttcacaaaac acactcgcca
     6241 accccgagcg catagttcaa aaccggagct tcagctactt aagaagatag gtacataaaa
     6301 ccgaccaaag aaactgacgc ctcacttatc cctcccctca ccagaggtcc ggcgcctgtc
     6361 gattcaggag agcctaccct aggcccgaac cctgcgtcct gcgacggaga aaagcctacc
     6421 gcacacctac cggctggtgg ccccaccctg cattataagc caacagaacg ggtgacgtca
     6481 cgacacgacg agggcgcgcg ctcccaaagg tacgggtgca ctgcccaacg gcaccgccat
     6541 aactgccgcc cccgcaacag acgacaaacc gagttctcca gtcagtgaca aacttcacgt
     6601 cagggtcccc agatggtgcc cagcccatct cacccgaata agagctttcc cgca
//
