LOCUS       sequence-312896-        7034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7034)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7034)
  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-m4133932
FEATURES             Location/Qualifiers
     source          1..7034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1456..1476
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1477..1521
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1574..1695
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1585..1604)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1639..1658
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1810..1856
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1886..2022
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2067..2138
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2364..2952)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2444..2463)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3452..4661
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4498..4517
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4665..5264
                     /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(4665..4684)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5161..5181
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5291..5339
                     /note="synthetic polyadenylation signal"
     CDS             join(5605..5673,5674..6465)
                     /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(5823..5842)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6478..6614
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6766..6837
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6842..6888)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6906..6954
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgagcgc cgttaagcgg
      601 acaatcaaag ggacccatca ctggttgctc cttactatac tgacgtcttt actggtcttg
      661 gtacagagta cacaatggtc gcttttcttt tttctgtacg agaatgcatt cctcccattt
      721 gcgatgggaa tcattgctat gtccgccttc gctatgatgt tcgtgaagca caaacatgcc
      781 ttcctttgcc tgtttctcct accctcactg gcaaccgtgg cctattttaa catggtgtac
      841 atgcctgcga gctgggtcat gcgaattatg acttggctgg acatggttga tacctcccta
      901 tctggcttca agttaaagga ctgtgtgatg tatgcatcag ctgtagtgct gctaatcctg
      961 atgacagctc gcacagtcta cgatgacggt gccaggagag tgtggactct gatgaatgtc
     1021 ctcaccttag tttataaagt atactatggc aacgctctcg atcaggccat tagtatgtgg
     1081 gcattgataa tctccgtgac gagtaactac agcggagtgg taaccactgt catgtttttg
     1141 gcacgcggga tagtgttcat gtgtgtcgaa tattgcccga tcttttttat cacagggaat
     1201 actctgcaat gcattatgct ggtttactgt ttcttgggct atttctgcac ctgttatttt
     1261 ggtctattct gcctgcttaa taggtacttt cgtttgacac taggagtcta tgattacttg
     1321 gtgagcacgc aggagtttcg gtatatgaac agtcagggcc tgcttccccc aaagaactct
     1381 attgacgcgt ttaaactcaa tataaagctg ttgggcgtgg gtggaaaacc ttgcatcaag
     1441 gtggccactg tgcaggaaaa cctttacttt cagtcgaaag aaaccgcggc agctaagttc
     1501 gagaggcagc acatggacag ttaagctagc ttgactgact gagatacagc gtaccttcag
     1561 ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     1621 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     1681 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     1741 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg cccatctcta
     1801 tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtgcc
     1861 cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct
     1921 gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc
     1981 ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca gctatcctgc
     2041 aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag
     2101 gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct
     2161 atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc
     2221 gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct
     2281 ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2341 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2401 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2461 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2521 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2581 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2641 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2701 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2761 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2821 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2881 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2941 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3001 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3061 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3121 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3181 atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt
     3241 cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc
     3301 ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat
     3361 ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag
     3421 atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc
     3481 gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg caggagcgtc
     3541 ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag
     3601 aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg
     3661 ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg
     3721 agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac
     3781 agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc
     3841 gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct
     3901 cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag
     3961 gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg
     4021 aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg
     4081 ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg ggaaagctct tattcgggtg
     4141 agatgggctg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac tggagaactc
     4201 ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag tgcacccgta
     4261 cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg
     4321 cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg
     4381 gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg
     4441 gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt aagtagctga
     4501 agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa gttttttagg
     4561 caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag acttgtaaat
     4621 tgtccgctaa attctggccg tttttggctt ttttgttaga caacatgacc gagtacaagc
     4681 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     4741 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     4801 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     4861 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     4921 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     4981 agcaacagat ggagggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     5041 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     5101 ccggagtgga ggccgccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     5161 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     5221 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctaggctagt atgtaagcct
     5281 agtcttagat aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt
     5341 gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc
     5401 catatcgttg cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc
     5461 gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat
     5521 tgatagtctg atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag
     5581 acaggatcag caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc
     5641 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5701 gatgctgaag atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt
     5761 aagatccttg agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt
     5821 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5881 atacactatt ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg
     5941 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6001 gccaacttac ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac
     6061 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6121 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta
     6181 actggcgaac tacttactct agcttcccgg caacagttga tagactggat ggaggcggat
     6241 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6301 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6361 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6421 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca
     6481 ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag
     6541 attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag
     6601 aaatttatcc ttaagatccc gaatcgttta aactcgactc tggctctatc gaatctccgt
     6661 cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt
     6721 cagctatcgt cagcttacct ttttggcagc gatcgcggct cccgacatct tggaccatta
     6781 gctccacagg tatcttcttc cctctagtgg tcataacagc agcttcagct acctctcaat
     6841 tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctatc aatcgttgcg
     6901 ttacacacac aaaaaaccaa cacacatcca tcttcgatgg atagcgattt tattatctaa
     6961 ctgctgatcg agtgtagcca gatctagtaa tcaattacgg ggtcattagt tcatagccca
     7021 tatatggagt tccg
//
