LOCUS       sequence-300696-        6248 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 6248)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6248)
  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-m4252804
FEATURES             Location/Qualifiers
     source          1..6248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      104..113
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             224..244
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             245..262
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    315..436
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(326..345)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     380..399
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      551..597
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      627..763
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    808..879
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1105..1693)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1185..1204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2193..3402
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3239..3258
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3406..4005
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3406..3425)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3902..3922
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4032..4080
                     /note="synthetic polyadenylation signal"
     CDS             join(4346..4414,4415..5206)
                     /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(4564..4583)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5219..5355
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5507..5578
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5583..5629)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5647..5695
                     /note="synthetic polyadenylation signal"
     enhancer        5776..6079
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6081..6248,1..35)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(6234..6248,1..6)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat
       61 ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgccacca tggggtacat
      121 aaatgttttc gcctttccat tcacgatcta ttcactcctt ttatgtagga tgaactctcg
      181 caattacatt gctcaggtag acgtcgtgaa ctttaaccta actgaaaacc tttactttca
      241 gtcgcaccat caccatcacc attaagctag cttgactgac tgagatacag cgtaccttca
      301 gctcacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
      361 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
      421 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
      481 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct
      541 atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc
      601 ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc
      661 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
      721 cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg
      781 caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta
      841 ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc
      901 tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt
      961 cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc
     1021 tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1081 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1141 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1201 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1261 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1321 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1381 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1441 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1501 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1561 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1621 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1681 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1741 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     1801 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     1861 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     1921 catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt
     1981 tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc
     2041 cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata
     2101 tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga
     2161 gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc
     2221 cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt
     2281 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta
     2341 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg
     2401 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg
     2461 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca
     2521 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat
     2581 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc
     2641 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa
     2701 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt
     2761 gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt
     2821 gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt
     2881 gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact
     2941 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt
     3001 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat
     3061 gcagggtggg gccaccagcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag
     3121 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag
     3181 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg
     3241 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag
     3301 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa
     3361 ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag
     3421 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     3481 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     3541 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     3601 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     3661 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     3721 cagcaacaga tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     3781 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     3841 cccggagtgg aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     3901 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     3961 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc
     4021 tagtcttaga taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg
     4081 tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat
     4141 ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat
     4201 cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata
     4261 ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga
     4321 gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc
     4381 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4441 agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg
     4501 taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt
     4561 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4621 catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac
     4681 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4741 ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa
     4801 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4861 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt
     4921 aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga
     4981 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5041 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5101 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5161 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc
     5221 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     5281 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     5341 gaaatttatc cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg
     5401 tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg
     5461 tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt
     5521 agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa
     5581 ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc
     5641 gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta
     5701 actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc
     5761 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     5821 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     5881 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     5941 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     6001 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     6061 agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     6121 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     6181 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     6241 gggcggta
//
