LOCUS       sequence-320319-        4401 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 4401)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4401)
  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-m4134393
FEATURES             Location/Qualifiers
     source          1..4401
                     /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             718..795
                     /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"
     polyA_signal    848..969
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(859..878)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     913..932
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1084..1130
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1160..1296
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1341..1412
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1638..2226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1718..1737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2972..3040,3041..3832)
                     /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(3190..3209)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3845..3981
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4133..4204
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4209..4255)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4273..4321
                     /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 ccatggggta cataaatgtt
      601 ttcgcctttc cattcacgat ctattcactc cttttatgta ggatgaactc tcgcaattac
      661 attgctcagg tagacgtcgt gaactttaac ctaactgaaa acctttactt tcagtcgaag
      721 aggcgatgga aaaagaattt tatcgcggtg agtgcagcca acagattcaa aaagatatcc
      781 tcgtctggtg ccctctaagc tagcttgact gactgagata cagcgtacct tcagctcaca
      841 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
      901 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
      961 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     1021 gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta
     1081 tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg
     1141 gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     1201 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     1261 ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcaggcga
     1321 tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca
     1381 gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg ggctatcgaa
     1441 acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac
     1501 gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt
     1561 acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1621 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     1681 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1741 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1801 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     1861 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     1921 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     1981 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2041 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2101 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2161 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2221 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2281 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2341 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2401 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2461 gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg
     2521 atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc
     2581 cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga atatccgaga
     2641 tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc cgagatccga
     2701 ggaatatcga aatcggggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc
     2761 gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag
     2821 tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc
     2881 tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta
     2941 tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc gtgtcgccct
     3001 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     3061 agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa
     3121 cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt
     3181 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     3241 tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca
     3301 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     3361 cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt
     3421 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     3481 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa
     3541 actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga
     3601 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3661 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3721 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3781 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct
     3841 aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     3901 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     3961 ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa
     4021 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
     4081 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
     4141 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
     4201 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat
     4261 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat
     4321 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca
     4381 tagcccatat atggagttcc g
//
