LOCUS       sequence-300244-        6785 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 6785)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6785)
  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-m4133534
FEATURES             Location/Qualifiers
     source          1..6785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             697..717
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             751..828
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             856..876
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1325..1446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1336..1355)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1390..1409
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1561..1607
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1637..1773
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1818..1889
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2115..2703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2195..2214)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3203..4412
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4249..4268
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4416..5015
                     /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(4416..4435)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4912..4932
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5042..5090
                     /note="synthetic polyadenylation signal"
     CDS             join(5356..5424,5425..6216)
                     /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(5574..5593)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6229..6365
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6517..6588
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6593..6639)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6657..6705
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgggcta tataaacgtt
      601 ttcgcttttc cgtttacgat atatagtcta ctcttgtgca gaatgaattc tcgtaactac
      661 atagcacaag tagatgtagt taactttaat ctcacagaaa acctttactt tcagtcggga
      721 cggcgaatcc cggggctgat aaatccctgg aaaaggagat ggaagaagaa ctttattgca
      781 gtgtctgcgg ctaaccggtt caaaaagatc agttcaagcg gcgccctaga ttacgacatt
      841 cctacgaccg cctccgagaa tctctatttc cagggtgaat ttggcttggc acaacatgac
      901 gaggccgtcg ataacaagtt taataaagaa cagcagaacg ctttctatga gatccttcac
      961 ttaccaaatc tgaacgaaga gcaacgcaac gctttcattc agagcctgaa agacgatccc
     1021 tcgcagtctg caaatctcct ggccgaagcc aagaagctga atgacgcaca ggctccaaaa
     1081 gttgataaca agtttaacaa ggagcagcag aatgctttct acgagatatt gcaccttcct
     1141 aatctgaatg aggaacaaag aaatgcattt atccagtccc taaaagacga tccatcacag
     1201 agtgccaacc tgcttgcgga agccaaaaag ctcaatgatg cccaggcccc gaaagtagac
     1261 gcgaaccatc aataagctag cttgactgac tgagatacag cgtaccttca gctcacagac
     1321 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     1381 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     1441 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     1501 gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg
     1561 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc
     1621 ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     1681 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     1741 tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct
     1801 ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa
     1861 gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact
     1921 taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct
     1981 cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg
     2041 agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2101 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2161 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2221 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2281 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2341 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2401 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2461 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2521 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2581 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2641 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2701 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2761 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2821 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2881 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca
     2941 tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc
     3001 catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt
     3061 ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg
     3121 ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga
     3181 atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc
     3241 cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt
     3301 ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt
     3361 atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc
     3421 cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc
     3481 cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc
     3541 cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg
     3601 tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac
     3661 ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg
     3721 aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga
     3781 cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag
     3841 aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct
     3901 gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg
     3961 tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga
     4021 gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg
     4081 gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc
     4141 tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga
     4201 ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt
     4261 tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg
     4321 aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta
     4381 aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag cccacggtgc
     4441 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     4501 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     4561 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     4621 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     4681 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     4741 tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     4801 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     4861 aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     4921 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     4981 gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc tagtcttaga
     5041 taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta
     5101 ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt
     5161 gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat
     5221 tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct
     5281 gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca
     5341 gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5401 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5461 gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5521 gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt
     5581 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5641 tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg
     5701 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5761 cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5821 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5881 cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa
     5941 ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca
     6001 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6061 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6121 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6181 gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag
     6241 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
     6301 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
     6361 cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag
     6421 cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg
     6481 tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag
     6541 gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac
     6601 ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca
     6661 caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc
     6721 gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc atatatggag
     6781 ttccg
//
