LOCUS       sequence-295070-        4713 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 4713)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4713)
  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-m4133967
FEATURES             Location/Qualifiers
     source          1..4713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             119..139
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             488..505
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    582..703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(593..612)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     647..666
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      818..864
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      894..1030
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1075..1146
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1372..1960)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1452..1471)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2706..2774,2775..3566)
                     /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(2924..2943)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3579..3715
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3867..3938
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3943..3989)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4007..4055
                     /note="synthetic polyadenylation signal"
     enhancer        4136..4439
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4441..4643
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4594..4614
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4712..4713,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatgggc tatataaacg ttttcgcttt tccgtttacg atatatagtc tactcttgtg
       61 cagaatgaat tctcgtaact acatagcaca agtagatgta gttaacttta atctcacaga
      121 aaacctttac tttcagtcga tgagtgacaa aatcattcat ctaaccgatg actcctttga
      181 tacagacgtg ttgaaggccg atggtgcgat actggtagat ttctgggctg aatggtgtgg
      241 cccgtgcaag atgatcgcac ccattctgga cgagatcgcc gacgagtatc aaggaaaatt
      301 aactgtcgct aagcttaata tagatcagaa cccagggacg gcacctaaat acggcatcag
      361 gggaatacct accctcctgc tctttaagaa tggggaagtt gccgcgacaa aagtgggagc
      421 cctgtctaag ggccagctaa aagagttctt ggacgctaac ctggcaggtt caggctcggg
      481 gcacatgcac catcaccatc accatagcag cgggctcgtg ccacgcggct aagctagctt
      541 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt
      601 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
      661 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
      721 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
      781 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct
      841 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg
      901 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
      961 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     1021 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc
     1081 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc
     1141 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta
     1201 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg
     1261 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc
     1321 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1381 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1441 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1501 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1561 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1621 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1681 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1741 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1801 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1861 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1921 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1981 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2041 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2101 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2161 ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag
     2221 gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa
     2281 cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat
     2341 cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac
     2401 ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg
     2461 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt
     2521 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag
     2581 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa
     2641 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct
     2701 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2761 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2821 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2881 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2941 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3001 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3061 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3121 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3181 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3241 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc
     3301 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc
     3361 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3421 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3481 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3541 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc
     3601 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     3661 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc
     3721 cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa
     3781 cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc
     3841 tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt
     3901 ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac
     3961 ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca
     4021 acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc
     4081 agatctagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     4141 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     4201 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     4261 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     4321 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     4381 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
     4441 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     4501 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     4561 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     4621 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc
     4681 taccatccac tcgacacacc cgccagcggc cgc
//
