LOCUS       sequence-307353-        6368 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 6368)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6368)
  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-m4252823
FEATURES             Location/Qualifiers
     source          1..6368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    88..136
                     /note="synthetic polyadenylation signal"
     CDS             join(402..470,471..1262)
                     /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(620..639)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      1275..1411
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1563..1634
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1639..1685)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    1703..1751
                     /note="synthetic polyadenylation signal"
     enhancer        1832..2135
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2137..2339
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2290..2310
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2597..2617
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2621..2686
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2739..2860
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2750..2769)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2804..2823
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2975..3021
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3051..3187
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3232..3303
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3529..4117)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3609..3628)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4617..5826
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5663..5682
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(5830..6368,1..61)
                     /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(5830..5849)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6326..6346
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta
       61 ggctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt
      121 gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag
      181 tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg
      241 acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc
      301 tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg
      361 caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc
      421 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
      481 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac
      541 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga
      601 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
      661 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca
      721 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
      781 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa
      841 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
      901 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
      961 ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag
     1021 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     1081 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     1141 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     1201 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     1261 aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     1321 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     1381 cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg
     1441 ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt
     1501 cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc
     1561 gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc
     1621 ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta
     1681 tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata
     1741 gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt
     1801 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     1861 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     1921 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     1981 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     2041 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     2101 agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca
     2161 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     2221 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     2281 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     2341 gtttagtgaa ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc
     2401 agcggccgcc accatgttcc acttggtgga ttttcaggtc acgatcgcag aaatactgct
     2461 aattatcatg cgaaccttca aggttagcat ttggaacctg gactacatca ttaatctcat
     2521 cataaaaaac cttagtaagt ctttaactga gaataaatat tcccaactgg atgaggaaca
     2581 gccaatggag attgacgaaa acctttactt tcagtcgatg gattataagg accacgatgg
     2641 agactacaaa gatcatgaca ttgattacaa ggacgacgat gacaaataag ctagcttgac
     2701 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
     2761 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2821 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2881 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2941 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
     3001 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
     3061 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     3121 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     3181 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
     3241 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
     3301 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
     3361 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     3421 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     3481 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3541 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3601 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3661 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3721 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3781 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3841 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3901 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3961 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4021 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4081 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4141 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4201 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4261 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4321 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     4381 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     4441 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     4501 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     4561 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
     4621 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     4681 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     4741 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     4801 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     4861 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     4921 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     4981 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     5041 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     5101 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     5161 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     5221 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     5281 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     5341 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     5401 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     5461 tcacccgttc tgttggctta taatgcaggg tggggccacc agccggtagg tgtgcggtag
     5521 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     5581 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     5641 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     5701 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     5761 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     5821 ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     5881 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     5941 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     6001 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     6061 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     6121 ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg
     6181 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     6241 ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc
     6301 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     6361 gtcaccgc
//
