LOCUS       sequence-297419-        4368 bp    DNA     circular SYN 15-APR-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 4368)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4368)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4133528
FEATURES             Location/Qualifiers
     source          1..4368
                     /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             140..184
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    237..358
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(248..267)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     302..321
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      473..519
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      549..685
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    730..801
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1027..1615)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1107..1126)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2361..2429,2430..3221)
                     /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(2579..2598)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3234..3370
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3522..3593
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3598..3644)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3662..3710
                     /note="synthetic polyadenylation signal"
     enhancer        3791..4094
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4096..4298
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4249..4269
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4367..4368,1..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 aagaaaccgc ggcagctaag ttcgagaggc agcacatgga
      181 cagttaagct agcttgactg actgagatac agcgtacctt cagctcacag acatgataag
      241 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
      301 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
      361 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
      421 aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa
      481 ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct
      541 atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
      601 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
      661 ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt
      721 cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca
      781 tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac
      841 cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg
      901 cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat
      961 ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1021 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1081 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1141 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1201 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1261 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1321 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1381 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1441 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1501 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1561 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1621 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1681 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1741 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1801 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt
     1861 tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc
     1921 aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg
     1981 cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc
     2041 cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa
     2101 atcggggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     2161 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     2221 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     2281 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     2341 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     2401 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2461 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2521 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     2581 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     2641 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     2701 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2761 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2821 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2881 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     2941 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     3001 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3061 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3121 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3181 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     3241 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     3301 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     3361 ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt
     3421 tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc
     3481 tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc
     3541 cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa
     3601 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac
     3661 acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc
     3721 tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata
     3781 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     3841 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     3901 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     3961 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     4021 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     4081 tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     4141 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     4201 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     4261 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca
     4321 gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgc
//
