LOCUS       sequence-297524-        4392 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 4392)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4392)
  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-m4133390
FEATURES             Location/Qualifiers
     source          1..4392
                     /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             143..208
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    261..382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(272..291)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     326..345
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      497..543
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      573..709
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    754..825
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1051..1639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1131..1150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2385..2453,2454..3245)
                     /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(2603..2622)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3258..3394
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3546..3617
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3622..3668)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3686..3734
                     /note="synthetic polyadenylation signal"
     enhancer        3815..4118
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4120..4322
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4273..4293
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4391..4392,1..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 tggattataa ggaccacgat ggagactaca aagatcatga
      181 cattgattac aaggacgacg atgacaaata agctagcttg actgactgag atacagcgta
      241 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
      301 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
      361 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
      421 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc
      481 atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
      541 ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg
      601 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
      661 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct
      721 atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca
      781 ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc
      841 ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct
      901 gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac
      961 ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag
     1021 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1081 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1141 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1201 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1261 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1321 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1381 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1441 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1501 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1561 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1621 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1681 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1741 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1801 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1861 ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca
     1921 aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg
     1981 gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc
     2041 cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct
     2101 atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg tgtaccgaga acgatcctct
     2161 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     2221 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     2281 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     2341 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     2401 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2461 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     2521 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     2581 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2641 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     2701 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2761 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     2821 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2881 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2941 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     3001 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3061 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3121 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3181 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3241 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     3301 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     3361 atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc
     3421 tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct
     3481 cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct
     3541 cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc
     3601 agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     3661 ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg
     3721 atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg
     3781 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     3841 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     3901 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     3961 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     4021 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     4081 ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca
     4141 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     4201 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     4261 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     4321 ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc
     4381 gccagcggcc gc
//
