LOCUS       sequence-300727-        4341 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 4341)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4341)
  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-m4134504
FEATURES             Location/Qualifiers
     source          1..4341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    75..123
                     /note="synthetic polyadenylation signal"
     terminator      141..187
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    192..263
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(415..551)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(564..1355,1356..1424))
                     /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     1187..1206
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2170..2758
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2659..2678
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2984..3055
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3100..3236)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(3266..3312)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3427..3548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3464..3483)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3518..3537
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3601..3618)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3619..3639)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      3750..3759
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3828..4030)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3857..3877)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4032..4335
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tccatatatg ggctatgaac taatgacccc gtaattgatt actagatctg gctacactcg
       61 atcagcagtt agataataaa atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt
      121 gtgtgtaacg caacgattga tagcataacc ccttggggcc tctaaacggg tcttgagggg
      181 ttttttgaat tgagaggtag ctgaagctgc tgttatgacc actagaggga agaagatacc
      241 tgtggagcta atggtccaag atgtcgggag ccgcgatcgc tgccaaaaag gtaagctgac
      301 gatagctgac gagattcgat gcccgacgag caaatgagcg ccacggctgt tcgcgtaagc
      361 tcgaaacgac ggagattcga tagagccaga gtcgagttta aacgattcgg gatcttaagg
      421 ataaatttct ggtaaggagg acacgtatgg aagtgggcaa gttggggaag ccgtatccgt
      481 tgctgaatct ggcatatgtg ggagtataag acgcgcagcg tcgcatcagg catttttttc
      541 tgcgccaatg cacctagaat cggttaccaa tgcttaatca gtgaggcacc tatctcagcg
      601 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
      661 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
      721 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
      781 gcaactttat ccgcctccat ccagtctatc aactgttgcc gggaagctag agtaagtagt
      841 tcgccagtta atagtttacg caaggttgtt gccattgcta caggcatcgt ggtgtcacgc
      901 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
      961 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccaatcgt tgtcagaagt
     1021 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     1081 atgccatccg taagatgctt ttctgtgact ggtgaatact caaccaagtc attctgagaa
     1141 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     1201 catagcagaa ctttaaaagt gctcatcatt ggaaagcgtt cttcggggcg aaaactctca
     1261 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgcgcacc caactgatct
     1321 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     1381 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatgcgaaa gcctcctgct
     1441 gatcctgtct cttgatagat gtttgcaaaa gctaggactc gattatacgt tgaccgatca
     1501 gactatcaat atctaggttt aaacagatcg gtacgctgcc gtgaccaggc ttgagtacaa
     1561 tatctgacga ttggacttcc tgggtcccaa gctggattcc gactggctga ctgccaagca
     1621 acgatatgga tcgtcgagac tcgcactgag aggatcgttc tcggtacacc aggcgcgccc
     1681 cgatttcgat attcctcgga tctcggatag atcgggattg aggatgtagg ttgaacttct
     1741 ctcgggtgat cccgatctcg gatattcggg attggagtaa tacctggcga taggcgctgc
     1801 caagcaatag ccaaggcgca aggccggcca agagacttgc gatagttcgt tcgagaggta
     1861 gcctgcaaga tggatcggac gaaaggtttg aagattttcc gacgagggcc ttggagagtt
     1921 cggaaattta aatgcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1981 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2041 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2101 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2161 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2221 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2281 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     2341 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2401 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2461 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2521 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2581 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2641 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2701 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2761 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgat
     2821 ttaaatgaaa ctcgtaatcg gagaacgggt gcaatagcaa ggaccagatc gaacttgact
     2881 gacgcaattc tgagcgtagt cgaaacccag acaagtcgat ctcgccaagt agctgacttg
     2941 actggttaat taagtttcga tagcccaagg taaccaatga gattagggag ggaagaaagt
     3001 ttgatgaact acttctgacc cctagtggtg tccagaaaag accattaaag gaatgtcttg
     3061 atcgaaatcg agagatcgcc tgcaggatag ctgacgacct taaggataaa tttctggtaa
     3121 ggaggacacg tatggaagtg ggcaagttgg ggaagccgta tccgttgctg aatctggcat
     3181 atgtgggagt ataagacgcg cagcgtcgca tcaggcattt ttttctgcgc caatgccggt
     3241 agatagcaat ccggcccgag gggcacaaaa aacccctcaa gacccgttta gaggccccaa
     3301 ggggttatgc tacgataccg atagagatgg gccaatacca catttgtaga ggttttactt
     3361 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3421 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3481 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3541 gtatcttatc atgtctgtga gctgaaggta cgctgtatct cagtcagtca agctagctta
     3601 atggtgatgg tgatggtgcg actgaaagta aaggttttca gttaggttaa agttcacgac
     3661 gtctacctga gcaatgtaat tgcgagagtt catcctacat aaaaggagtg aatagatcgt
     3721 gaatggaaag gcgaaaacat ttatgtaccc catggtggcg gccgctggcg ggtgtgtcga
     3781 gtggatggta ggatcgacaa agatctgatc tgacggttca ctaaaccagc tctgcttata
     3841 tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga
     3901 cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg
     3961 tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa
     4021 aaccgcatca gcatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag
     4081 tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca
     4141 atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt
     4201 aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac
     4261 gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc
     4321 gtaagttatg taacgcggaa c
//
