LOCUS       sequence-295968-        7060 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 7060)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7060)
  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-m4133724
FEATURES             Location/Qualifiers
     source          1..7060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             878..898
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             899..976
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    1029..1150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1040..1059)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1094..1113
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1265..1311
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1341..1477
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1522..1593
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1819..2407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1899..1918)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2907..4116
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3953..3972
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4120..4719
                     /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(4120..4139)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4616..4636
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4746..4794
                     /note="synthetic polyadenylation signal"
     CDS             join(5060..5128,5129..5920)
                     /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(5278..5297)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5933..6069
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6214..6285
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6290..6336)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6354..6402
                     /note="synthetic polyadenylation signal"
     enhancer        6483..6786
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6788..6990
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6941..6961
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgagc gccgttaagc ggacaatcaa agggacccat cactggttgc tccttactat
       61 actgacgtct ttactggtct tggtacagag tacacaatgg tcgcttttct tttttctgta
      121 cgagaatgca ttcctcccat ttgcgatggg aatcattgct atgtccgcct tcgctatgat
      181 gttcgtgaag cacaaacatg ccttcctttg cctgtttctc ctaccctcac tggcaaccgt
      241 ggcctatttt aacatggtgt acatgcctgc gagctgggtc atgcgaatta tgacttggct
      301 ggacatggtt gatacctccc tatctggctt caagttaaag gactgtgtga tgtatgcatc
      361 agctgtagtg ctgctaatcc tgatgacagc tcgcacagtc tacgatgacg gtgccaggag
      421 agtgtggact ctgatgaatg tcctcacctt agtttataaa gtatactatg gcaacgctct
      481 cgatcaggcc attagtatgt gggcattgat aatctccgtg acgagtaact acagcggagt
      541 ggtaaccact gtcatgtttt tggcacgcgg gatagtgttc atgtgtgtcg aatattgccc
      601 gatctttttt atcacaggga atactctgca atgcattatg ctggtttact gtttcttggg
      661 ctatttctgc acctgttatt ttggtctatt ctgcctgctt aataggtact ttcgtttgac
      721 actaggagtc tatgattact tggtgagcac gcaggagttt cggtatatga acagtcaggg
      781 cctgcttccc ccaaagaact ctattgacgc gtttaaactc aatataaagc tgttgggcgt
      841 gggtggaaaa ccttgcatca aggtggccac tgtgcaggaa aacctttact ttcagtcgaa
      901 gaggcgatgg aaaaagaatt ttatcgcggt gagtgcagcc aacagattca aaaagatatc
      961 ctcgtctggt gccctctaag ctagcttgac tgactgagat acagcgtacc ttcagctcac
     1021 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     1081 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     1141 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     1201 ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt
     1261 atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg
     1321 ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     1381 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     1441 tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg
     1501 atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc
     1561 agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga
     1621 aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta
     1681 cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat
     1741 tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1801 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1861 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1921 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1981 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2041 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2101 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2161 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2221 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2281 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2341 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2401 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2461 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2521 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2581 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2641 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc
     2701 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg
     2761 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag
     2821 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg
     2881 aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg
     2941 cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat
     3001 ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc
     3061 cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc
     3121 tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga
     3181 tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta
     3241 gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac
     3301 ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg
     3361 ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc
     3421 cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg
     3481 aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg
     3541 caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc gggtgagatg
     3601 ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt
     3661 gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt
     3721 gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg
     3781 tggggccacc tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg
     3841 ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa
     3901 gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc
     3961 cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct
     4021 tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg taaattgtcc
     4081 gctaaattct ggccgttttt ggcttttttg ttagacaaca tgaccgagta caagcccacg
     4141 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     4201 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     4261 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     4321 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     4381 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     4441 cagatggagg gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     4501 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     4561 gtggaggccg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     4621 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     4681 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctagg ctagtatgta agcctagtct
     4741 tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac
     4801 gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     4861 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     4921 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     4981 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     5041 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     5101 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5161 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5221 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     5281 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5341 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     5401 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5461 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5521 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5581 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     5641 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     5701 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5761 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5821 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5881 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     5941 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     6001 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     6061 tatccttaag atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt
     6121 acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca
     6181 gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta
     6241 tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc
     6301 tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa
     6361 aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag
     6421 tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     6481 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     6541 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     6601 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     6661 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     6721 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     6781 accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     6841 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     6901 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     6961 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg
     7021 tcgatcctac catccactcg acacacccgc cagcggccgc
//
