LOCUS       sequence-300707-        6272 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 6272)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6272)
  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-m4252800
FEATURES             Location/Qualifiers
     source          1..6272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    47..95
                     /note="synthetic polyadenylation signal"
     terminator      113..159
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    164..235
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(387..523)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(536..1327,1328..1396))
                     /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     1159..1178
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1662..1710
                     /note="synthetic polyadenylation signal"
     CDS             complement(1737..2336)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1820..1840)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2317..2336
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2340..3549)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2484..2503)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4049..4637
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4538..4557
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4863..4934
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4979..5115)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5145..5191)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5306..5427
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5343..5362)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5397..5416
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5480..5521)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(5522..5542)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      5653..5662
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5731..5933)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5760..5780)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5935..6238
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ccgtaattga ttactagatc tggctacact cgatcagcag ttagataata aaatcgctat
       61 ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgcaacgatt gatagcataa
      121 ccccttgggg cctctaaacg ggtcttgagg ggttttttga attgagaggt agctgaagct
      181 gctgttatga ccactagagg gaagaagata cctgtggagc taatggtcca agatgtcggg
      241 agccgcgatc gctgccaaaa aggtaagctg acgatagctg acgagattcg atgcccgacg
      301 agcaaatgag cgccacggct gttcgcgtaa gctcgaaacg acggagattc gatagagcca
      361 gagtcgagtt taaacgattc gggatcttaa ggataaattt ctggtaagga ggacacgtat
      421 ggaagtgggc aagttgggga agccgtatcc gttgctgaat ctggcatatg tgggagtata
      481 agacgcgcag cgtcgcatca ggcatttttt tctgcgccaa tgcacctaga atcggttacc
      541 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
      601 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
      661 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
      721 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
      781 tcaactgttg ccgggaagct agagtaagta gttcgccagt taatagttta cgcaaggttg
      841 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
      901 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
      961 gctccttcgg tcctccaatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     1021 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     1081 ctggtgaata ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     1141 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     1201 ttggaaagcg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     1261 cgatgtaacc cactcgcgca cccaactgat cttcagcatc ttttactttc accagcgttt
     1321 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     1381 aatgttgaat actcatgcga aagcctcctg ctgatcctgt ctcttgatag atgtttgcaa
     1441 aagctaggac tcgattatac gttgaccgat cagactatca atatctaggt ttaaacagat
     1501 cggtacgctg ccgtgaccag gcttgagtac aatatctgac gattggactt cctgggtccc
     1561 aagctggatt ccgactggct gactgccaag caacgatatg gatcgtcgag actcgcactg
     1621 agaggatcgt tctcggtaca ccaggcgcgc ctagcgttac acacacaaaa aaccaacaca
     1681 catccatctt cgatggatag cgattttatt atctaagact aggcttacat actagcctag
     1741 gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg
     1801 gtgacggtga agccgagccg ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg
     1861 aaggcgggca ccccggcgcg ctcggcggcc tccactccgg ggagcacgac ggcgctgccc
     1921 agacccttgc cctggtggtc gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc
     1981 ttgggccggt gcggcgccag gaggccctcc atctgttgct gcgcggccag ccgggaaccg
     2041 ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc
     2101 ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg
     2161 agcccgacgc gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc
     2221 ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg
     2281 gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatgttg
     2341 tctaacaaaa aagccaaaaa cggccagaat ttagcggaca atttacaagt ctaacactga
     2401 aaattacata ttgacccaaa tgattacatt tcaaaaggtg cctaaaaaac ttcacaaaac
     2461 acactcgcca accccgagcg catagttcaa aaccggagct tcagctactt aagaagatag
     2521 gtacataaaa ccgaccaaag aaactgacgc ctcacttatc cctcccctca ccagaggtcc
     2581 ggcgcctgtc gattcaggag agcctaccct aggcccgaac cctgcgtcct gcgacggaga
     2641 aaagcctacc gcacacctac cggctggtgg ccccaccctg cattataagc caacagaacg
     2701 ggtgacgtca cgacacgacg agggcgcgcg ctcccaaagg tacgggtgca ctgcccaacg
     2761 gcaccgccat aactgccgcc cccgcaacag acgacaaacc gagttctcca gtcagtgaca
     2821 aacttcacgt cagggtcccc agatggtgcc cagcccatct cacccgaata agagctttcc
     2881 cgcattagcg aagggctcaa gaccttgggt tcttgccgcc caccatgccc cccaccttgt
     2941 ttcaacgacc tcacagcccg cctcacaagc gtcttccatt caagactcgg gaacagccgc
     3001 cattttgctg cgctcccccc aacccccagt tcagggcaac cttgctcgcg gacccagact
     3061 acagcccttg gcggtctctc cacacgcttc cgtcccaccg agcggcccgg cggccacgaa
     3121 agccccggcc agcccagcag cccgctactc accaagtgac gatcacagcg atccacaaac
     3181 aagaaccgcg acccaaatcc cggctgcgac ggaactagct gtgccacacc cggcgcgtcc
     3241 ttatataatc atcggcgttc accgccccac ggagatccct ccgcagaatc gccgagaagg
     3301 gactactttt cctcgcctgt tccgctctct ggaaagaaaa ccagtgccct agagtcaccc
     3361 aagtcccgtc ctaaaatgtc cttctgctga tactggggtt ctaaggccga gtcttatgag
     3421 cagcgggccg ctgtcctgag cgtccgggcg gaaggatcag gacgctcctg cgcccttcgt
     3481 ctgacgtggc agcgctcgcc gtgacgaggg gggcgcccgc gggaggcgcc aaaacccggc
     3541 gcggaggcca ggcgcgcccc gatttcgata ttcctcggat ctcggataga tcgggattga
     3601 ggatgtaggt tgaacttctc tcgggtgatc ccgatctcgg atattcggga ttggagtaat
     3661 acctggcgat aggcgctgcc aagcaatagc caaggcgcaa ggccggccaa gagacttgcg
     3721 atagttcgtt cgagaggtag cctgcaagat ggatcggacg aaaggtttga agattttccg
     3781 acgagggcct tggagagttc ggaaatttaa atgcaactat ggatgaacga aatagacaga
     3841 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3901 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3961 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4021 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4081 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4141 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4201 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4261 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4321 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4381 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4441 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4501 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4561 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4621 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4681 ggccttttgc tcacatgatt taaatgaaac tcgtaatcgg agaacgggtg caatagcaag
     4741 gaccagatcg aacttgactg acgcaattct gagcgtagtc gaaacccaga caagtcgatc
     4801 tcgccaagta gctgacttga ctggttaatt aagtttcgat agcccaaggt aaccaatgag
     4861 attagggagg gaagaaagtt tgatgaacta cttctgaccc ctagtggtgt ccagaaaaga
     4921 ccattaaagg aatgtcttga tcgaaatcga gagatcgcct gcaggatagc tgacgacctt
     4981 aaggataaat ttctggtaag gaggacacgt atggaagtgg gcaagttggg gaagccgtat
     5041 ccgttgctga atctggcata tgtgggagta taagacgcgc agcgtcgcat caggcatttt
     5101 tttctgcgcc aatgccggta gatagcaatc cggcccgagg ggcacaaaaa acccctcaag
     5161 acccgtttag aggccccaag gggttatgct acgataccga tagagatggg ccaataccac
     5221 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     5281 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     5341 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5401 tttgtccaaa ctcatcaatg tatcttatca tgtctgtgag ctgaaggtac gctgtatctc
     5461 agtcagtcaa gctagcttag gtagaatcta agccgagaag tggattgggt ataggtttcc
     5521 ccgactgaaa gtaaaggttt tcagttaggt taaagttcac gacgtctacc tgagcaatgt
     5581 aattgcgaga gttcatccta cataaaagga gtgaatagat cgtgaatgga aaggcgaaaa
     5641 catttatgta ccccatggtg gcggccgctg gcgggtgtgt cgagtggatg gtaggatcga
     5701 caaagatctg atctgacggt tcactaaacc agctctgctt atatagacct cccaccgtac
     5761 acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg
     5821 ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     5881 ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcagcatggt
     5941 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     6001 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     6061 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     6121 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     6181 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg
     6241 gaactccata tatgggctat gaactaatga cc
//
