LOCUS       sequence-307356-        6689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF6 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6689)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6689)
  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-m4252805
FEATURES             Location/Qualifiers
     source          1..6689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..33
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(121..138)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(487..507)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(765..967)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(794..814)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        969..1272
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    1353..1401
                     /note="synthetic polyadenylation signal"
     terminator      1419..1465
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1470..1541
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1693..1829)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(1842..2633,2634..2702))
                     /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     2465..2484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    2968..3016
                     /note="synthetic polyadenylation signal"
     CDS             complement(3043..3642)
                     /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(3126..3146)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3623..3642
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3646..4855)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(3790..3809)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      5355..5943
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5844..5863
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6169..6240
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6285..6421)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(6451..6497)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    join(6612..6689,1..44)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6649..6668)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtgagctg
       61 aaggtacgct gtatctcagt cagtcaagct agcttagccg cgtggcacga gcccgctgct
      121 atggtgatgg tgatggtgca tgtgccccga gcctgaacct gccaggttag cgtccaagaa
      181 ctcttttagc tggcccttag acagggctcc cacttttgtc gcggcaactt ccccattctt
      241 aaagagcagg agggtaggta ttcccctgat gccgtattta ggtgccgtcc ctgggttctg
      301 atctatatta agcttagcga cagttaattt tccttgatac tcgtcggcga tctcgtccag
      361 aatgggtgcg atcatcttgc acgggccaca ccattcagcc cagaaatcta ccagtatcgc
      421 accatcggcc ttcaacacgt ctgtatcaaa ggagtcatcg gttagatgaa tgattttgtc
      481 actcatcgac tgaaagtaaa ggttttcatc aatctccatt ggttgctctt catctaattg
      541 agaatattta ttctcagtta gtgacttaga taaattttta attatgaggt ttatgatgta
      601 atcaagattc caaatggaaa ctttaaaagt cctcataata attagtaata tctctgctat
      661 agtaacctga aagtcaacga gatgaaacat ggtggcggcc gctggcgggt gtgtcgagtg
      721 gatggtagga tcgacaaaga tctgatctga cggttcacta aaccagctct gcttatatag
      781 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
      841 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
      901 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
      961 cgcatcagca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     1021 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     1081 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     1141 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
     1201 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     1261 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
     1321 tagatctggc tacactcgat cagcagttag ataataaaat cgctatccat cgaagatgga
     1381 tgtgtgttgg ttttttgtgt gtgtaacgca acgattgata gcataacccc ttggggcctc
     1441 taaacgggtc ttgaggggtt ttttgaattg agaggtagct gaagctgctg ttatgaccac
     1501 tagagggaag aagatacctg tggagctaat ggtccaagat gtcgggagcc gcgatcgctg
     1561 ccaaaaaggt aagctgacga tagctgacga gattcgatgc ccgacgagca aatgagcgcc
     1621 acggctgttc gcgtaagctc gaaacgacgg agattcgata gagccagagt cgagtttaaa
     1681 cgattcggga tcttaaggat aaatttctgg taaggaggac acgtatggaa gtgggcaagt
     1741 tggggaagcc gtatccgttg ctgaatctgg catatgtggg agtataagac gcgcagcgtc
     1801 gcatcaggca tttttttctg cgccaatgca cctagaatcg gttaccaatg cttaatcagt
     1861 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1921 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1981 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     2041 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctatcaa ctgttgccgg
     2101 gaagctagag taagtagttc gccagttaat agtttacgca aggttgttgc cattgctaca
     2161 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     2221 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     2281 ccaatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     2341 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgaatactca
     2401 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     2461 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaagcgttct
     2521 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     2581 cgcgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     2641 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     2701 atgcgaaagc ctcctgctga tcctgtctct tgatagatgt ttgcaaaagc taggactcga
     2761 ttatacgttg accgatcaga ctatcaatat ctaggtttaa acagatcggt acgctgccgt
     2821 gaccaggctt gagtacaata tctgacgatt ggacttcctg ggtcccaagc tggattccga
     2881 ctggctgact gccaagcaac gatatggatc gtcgagactc gcactgagag gatcgttctc
     2941 ggtacaccag gcgcgcctag cgttacacac acaaaaaacc aacacacatc catcttcgat
     3001 ggatagcgat tttattatct aagactaggc ttacatacta gcctaggcac cgggcttgcg
     3061 ggtcatgcac caggtgcgcg gtccttcggg cacctcgacg tcggcggtga cggtgaagcc
     3121 gagccgctcg tagaagggga ggttgcgggg cgcggaggtc tccaggaagg cgggcacccc
     3181 ggcgcgctcg gcggcctcca ctccggggag cacgacggcg ctgcccagac ccttgccctg
     3241 gtggtcgggc gagacgccga cggtggccag gaaccacgcg ggctccttgg gccggtgcgg
     3301 cgccaggagg ccctccatct gttgctgcgc ggccagccgg gaaccgctca actcggccat
     3361 gcgcgggccg atctcggcga acaccgcccc cgcttcgacg ctctccggcg tggtccagac
     3421 cgccaccgcg gcgccgtcgt ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt
     3481 gaggaagagt tcttgcagct cggtgacccg ctcgatgtgg cggtccggat cgacggtgtg
     3541 gcgcgtggcg gggtagtcgg cgaacgcggc ggcgagggtg cgtacggccc tggggacgtc
     3601 gtcgcgggtg gcgaggcgca ccgtgggctt gtactcggtc atgttgtcta acaaaaaagc
     3661 caaaaacggc cagaatttag cggacaattt acaagtctaa cactgaaaat tacatattga
     3721 cccaaatgat tacatttcaa aaggtgccta aaaaacttca caaaacacac tcgccaaccc
     3781 cgagcgcata gttcaaaacc ggagcttcag ctacttaaga agataggtac ataaaaccga
     3841 ccaaagaaac tgacgcctca cttatccctc ccctcaccag aggtccggcg cctgtcgatt
     3901 caggagagcc taccctaggc ccgaaccctg cgtcctgcga cggagaaaag cctaccgcac
     3961 acctaccggc tggtggcccc accctgcatt ataagccaac agaacgggtg acgtcacgac
     4021 acgacgaggg cgcgcgctcc caaaggtacg ggtgcactgc ccaacggcac cgccataact
     4081 gccgcccccg caacagacga caaaccgagt tctccagtca gtgacaaact tcacgtcagg
     4141 gtccccagat ggtgcccagc ccatctcacc cgaataagag ctttcccgca ttagcgaagg
     4201 gctcaagacc ttgggttctt gccgcccacc atgcccccca ccttgtttca acgacctcac
     4261 agcccgcctc acaagcgtct tccattcaag actcgggaac agccgccatt ttgctgcgct
     4321 ccccccaacc cccagttcag ggcaaccttg ctcgcggacc cagactacag cccttggcgg
     4381 tctctccaca cgcttccgtc ccaccgagcg gcccggcggc cacgaaagcc ccggccagcc
     4441 cagcagcccg ctactcacca agtgacgatc acagcgatcc acaaacaaga accgcgaccc
     4501 aaatcccggc tgcgacggaa ctagctgtgc cacacccggc gcgtccttat ataatcatcg
     4561 gcgttcaccg ccccacggag atccctccgc agaatcgccg agaagggact acttttcctc
     4621 gcctgttccg ctctctggaa agaaaaccag tgccctagag tcacccaagt cccgtcctaa
     4681 aatgtccttc tgctgatact ggggttctaa ggccgagtct tatgagcagc gggccgctgt
     4741 cctgagcgtc cgggcggaag gatcaggacg ctcctgcgcc cttcgtctga cgtggcagcg
     4801 ctcgccgtga cgaggggggc gcccgcggga ggcgccaaaa cccggcgcgg aggccaggcg
     4861 cgccccgatt tcgatattcc tcggatctcg gatagatcgg gattgaggat gtaggttgaa
     4921 cttctctcgg gtgatcccga tctcggatat tcgggattgg agtaatacct ggcgataggc
     4981 gctgccaagc aatagccaag gcgcaaggcc ggccaagaga cttgcgatag ttcgttcgag
     5041 aggtagcctg caagatggat cggacgaaag gtttgaagat tttccgacga gggccttgga
     5101 gagttcggaa atttaaatgc aactatggat gaacgaaata gacagatcgc tgagataggt
     5161 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5221 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5281 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5341 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5401 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5461 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5521 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5581 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5641 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5701 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5761 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5821 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5881 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5941 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6001 atgatttaaa tgaaactcgt aatcggagaa cgggtgcaat agcaaggacc agatcgaact
     6061 tgactgacgc aattctgagc gtagtcgaaa cccagacaag tcgatctcgc caagtagctg
     6121 acttgactgg ttaattaagt ttcgatagcc caaggtaacc aatgagatta gggagggaag
     6181 aaagtttgat gaactacttc tgacccctag tggtgtccag aaaagaccat taaaggaatg
     6241 tcttgatcga aatcgagaga tcgcctgcag gatagctgac gaccttaagg ataaatttct
     6301 ggtaaggagg acacgtatgg aagtgggcaa gttggggaag ccgtatccgt tgctgaatct
     6361 ggcatatgtg ggagtataag acgcgcagcg tcgcatcagg catttttttc tgcgccaatg
     6421 ccggtagata gcaatccggc ccgaggggca caaaaaaccc ctcaagaccc gtttagaggc
     6481 cccaaggggt tatgctacga taccgataga gatgggccaa taccacattt gtagaggttt
     6541 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     6601 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6661 caaatttcac aaataaagca tttttttca
//
