LOCUS       sequence-300731-        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"
     misc_feature    284..355
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(400..536)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(566..612)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    727..848
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(764..783)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     818..837
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(901..945)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(946..966)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      1077..1086
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1155..1357)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1184..1204)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1359..1662
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    1743..1791
                     /note="synthetic polyadenylation signal"
     terminator      1809..1855
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1860..1931
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2083..2219)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(2232..3023,3024..3092))
                     /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     2855..2874
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3838..4368,1..58)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4327..4346
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
       61 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgat
      121 ttaaatgaaa ctcgtaatcg gagaacgggt gcaatagcaa ggaccagatc gaacttgact
      181 gacgcaattc tgagcgtagt cgaaacccag acaagtcgat ctcgccaagt agctgacttg
      241 actggttaat taagtttcga tagcccaagg taaccaatga gattagggag ggaagaaagt
      301 ttgatgaact acttctgacc cctagtggtg tccagaaaag accattaaag gaatgtcttg
      361 atcgaaatcg agagatcgcc tgcaggatag ctgacgacct taaggataaa tttctggtaa
      421 ggaggacacg tatggaagtg ggcaagttgg ggaagccgta tccgttgctg aatctggcat
      481 atgtgggagt ataagacgcg cagcgtcgca tcaggcattt ttttctgcgc caatgccggt
      541 agatagcaat ccggcccgag gggcacaaaa aacccctcaa gacccgttta gaggccccaa
      601 ggggttatgc tacgataccg atagagatgg gccaatacca catttgtaga ggttttactt
      661 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
      721 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
      781 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
      841 gtatcttatc atgtctgtga gctgaaggta cgctgtatct cagtcagtca agctagctta
      901 actgtccatg tgctgcctct cgaacttagc tgccgcggtt tctttcgact gaaagtaaag
      961 gttttctgtg agattaaagt taactacatc tacttgtgct atgtagttac gagaattcat
     1021 tctgcacaag agtagactat atatcgtaaa cggaaaagcg aaaacgttta tatagcccat
     1081 ggtggcggcc gctggcgggt gtgtcgagtg gatggtagga tcgacaaaga tctgatctga
     1141 cggttcacta aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt
     1201 tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa
     1261 aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc
     1321 tatccacgcc cattgatgta ctgccaaaac cgcatcagca tggtaatagc gatgactaat
     1381 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
     1441 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
     1501 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     1561 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     1621 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     1681 ctatgaacta atgaccccgt aattgattac tagatctggc tacactcgat cagcagttag
     1741 ataataaaat cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgca
     1801 acgattgata gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgaattg
     1861 agaggtagct gaagctgctg ttatgaccac tagagggaag aagatacctg tggagctaat
     1921 ggtccaagat gtcgggagcc gcgatcgctg ccaaaaaggt aagctgacga tagctgacga
     1981 gattcgatgc ccgacgagca aatgagcgcc acggctgttc gcgtaagctc gaaacgacgg
     2041 agattcgata gagccagagt cgagtttaaa cgattcggga tcttaaggat aaatttctgg
     2101 taaggaggac acgtatggaa gtgggcaagt tggggaagcc gtatccgttg ctgaatctgg
     2161 catatgtggg agtataagac gcgcagcgtc gcatcaggca tttttttctg cgccaatgca
     2221 cctagaatcg gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2281 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2341 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2401 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2461 gcctccatcc agtctatcaa ctgttgccgg gaagctagag taagtagttc gccagttaat
     2521 agtttacgca aggttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2581 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2641 tgcaaaaaag cggttagctc cttcggtcct ccaatcgttg tcagaagtaa gttggccgca
     2701 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2761 agatgctttt ctgtgactgg tgaatactca accaagtcat tctgagaata gtgtatgcgg
     2821 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2881 ttaaaagtgc tcatcattgg aaagcgttct tcggggcgaa aactctcaag gatcttaccg
     2941 ctgttgagat ccagttcgat gtaacccact cgcgcaccca actgatcttc agcatctttt
     3001 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3061 ataagggcga cacggaaatg ttgaatactc atgcgaaagc ctcctgctga tcctgtctct
     3121 tgatagatgt ttgcaaaagc taggactcga ttatacgttg accgatcaga ctatcaatat
     3181 ctaggtttaa acagatcggt acgctgccgt gaccaggctt gagtacaata tctgacgatt
     3241 ggacttcctg ggtcccaagc tggattccga ctggctgact gccaagcaac gatatggatc
     3301 gtcgagactc gcactgagag gatcgttctc ggtacaccag gcgcgccccg atttcgatat
     3361 tcctcggatc tcggatagat cgggattgag gatgtaggtt gaacttctct cgggtgatcc
     3421 cgatctcgga tattcgggat tggagtaata cctggcgata ggcgctgcca agcaatagcc
     3481 aaggcgcaag gccggccaag agacttgcga tagttcgttc gagaggtagc ctgcaagatg
     3541 gatcggacga aaggtttgaa gattttccga cgagggcctt ggagagttcg gaaatttaaa
     3601 tgcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3661 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3721 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3781 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3841 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3901 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3961 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4021 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4081 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4141 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4201 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4261 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4321 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgcca
//
