LOCUS       sequence-297597-        6278 bp    DNA     circular SYN 15-APR-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 6278)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6278)
  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-m4252981
FEATURES             Location/Qualifiers
     source          1..6278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    240..361
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(251..270)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     305..324
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      476..522
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      552..688
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    733..804
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1030..1618)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1110..1129)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2118..3327
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3164..3183
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3331..3930
                     /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(3331..3350)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3827..3847
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3957..4005
                     /note="synthetic polyadenylation signal"
     CDS             join(4271..4339,4340..5131)
                     /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(4489..4508)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5144..5280
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5432..5503
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5508..5554)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5572..5620
                     /note="synthetic polyadenylation signal"
     enhancer        5701..6004
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6006..6208
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6159..6179
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgttc cacttggtgg attttcaggt cacgatcgca gaaatactgc taattatcat
       61 gcgaaccttc aaggttagca tttggaacct ggactacatc attaatctca tcataaaaaa
      121 ccttagtaag tctttaactg agaataaata ttcccaactg gatgaggaac agccaatgga
      181 gattgactaa gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat
      241 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
      301 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
      361 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
      421 ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca
      481 taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt
      541 gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
      601 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
      661 gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga
      721 tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt
      781 tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt
      841 aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa
      901 ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt
      961 catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1021 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1081 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1141 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1201 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1261 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1321 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1381 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1441 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1501 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1561 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1621 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1681 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1741 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1801 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa
     1861 atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct
     1921 tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta
     1981 ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc
     2041 acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc
     2101 gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
     2161 cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga tccttccgcc
     2221 cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag
     2281 cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag
     2341 agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg
     2401 ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg
     2461 cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt
     2521 agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag
     2581 cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg
     2641 ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt
     2701 gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc
     2761 aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat gggctgggca
     2821 ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt
     2881 tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg
     2941 cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac
     3001 cagccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg
     3061 taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt
     3121 cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga
     3181 actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg
     3241 taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc cgctaaattc
     3301 tggccgtttt tggctttttt gttagacaac atgaccgagt acaagcccac ggtgcgcctc
     3361 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     3421 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     3481 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     3541 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     3601 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggag
     3661 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     3721 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcc
     3781 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     3841 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     3901 tggtgcatga cccgcaagcc cggtgcctag gctagtatgt aagcctagtc ttagataata
     3961 aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc
     4021 gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata tcgttgcttg
     4081 gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca gatattgtac
     4141 tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat agtctgatcg
     4201 gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag gatcagcagg
     4261 aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4321 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4381 gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4441 ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4501 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4561 gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg gcatgacagt
     4621 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4681 gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4741 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4801 caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg gcgaactact
     4861 tactctagct tcccggcaac agttgataga ctggatggag gcggataaag ttgcaggacc
     4921 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4981 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5041 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5101 gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg cgcagaaaaa
     5161 aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat
     5221 acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa
     5281 gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac
     5341 gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc
     5401 ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc
     5461 ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc
     5521 aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa
     5581 aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg
     5641 tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     5701 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     5761 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     5821 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     5881 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     5941 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     6001 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     6061 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     6121 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     6181 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
     6241 gatcctacca tccactcgac acacccgcca gcggccgc
//
