LOCUS       sequence-295828-        5790 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5790)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5790)
  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-m4133483
FEATURES             Location/Qualifiers
     source          1..5790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1508..1528
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1529..1606
                     /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    1659..1780
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1670..1689)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1724..1743
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1895..1941
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1971..2107
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2152..2223
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2449..3037)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2529..2548)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3783..3851,3852..4643)
                     /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(4001..4020)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4656..4792
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4944..5015
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5020..5066)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5084..5132
                     /note="synthetic polyadenylation signal"
     enhancer        5213..5516
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5518..5720
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5671..5691
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaag attgtaaata actggttgaa acaactaata aaggttactt tagtgttcct
       61 gtttgtcgcc gcaatcttct atcttatcac cccggtgcac gtgatgagta aacatacaga
      121 cttttcttca gaaattatcg ggtacaaggc tatcgatggc ggtgtaacgc gcgacattgc
      181 ctccacagat acctgcttcg cgaacaaaca cgctgacttt gatacttggt tcagccagcg
      241 aggaggctcg tacaccaatg acaaagcctg tccactgatc gcagccgtga taactagaga
      301 ggttggattc gtcgtccctg ggctccccgg gacaattctc cgtacaacca acggcgactt
      361 tctgcatttt ctgccccggg tgttctccgc agtcggtaat atctgttata ctccatctaa
      421 gctcattgag tacacggatt ttgctaccag cgcgtgcgtg ttggccgcag aatgcaccat
      481 atttaaggac gcttctggaa aacctgtgcc ctattgttac gataccaatg tgcttgaggg
      541 ctcagttgcc tatgaaagtc tgaggccaga cactagatac gttttaatgg atggaagcat
      601 catccagttc ccaaacacgt atctggaggg gtccgtgcgg gtcgtgacca cattcgattc
      661 agaatactgc cgccacggta catgcgagag gtctgaggct ggcgtatgtg tgtcaactag
      721 cggtcgatgg gttctaaata acgattatta tcgttctctg cccggcgttt tttgtggagt
      781 ggacgccgtt aaccttttga ccaacatgtt tacacctctc attcagccca ttggtgcact
      841 ggatatctcc gcgagtattg tggctggggg aatagtcgcc attgtcgtaa cctgcttggc
      901 atactatttc atgcgtttca ggcgagcttt tggcgaatac tcacatgtcg tggccttcaa
      961 tactcttctg tttctgatga gcttcacggt actctgtctg accccagtat acagtttcct
     1021 acctggggtg tattctgtga tctacttata cctcacattt tacctgacca atgacgtgtc
     1081 ctttctggcc cacatccaat ggatggttat gttcacgccg ctcgtgcctt tctggattac
     1141 aatcgcttat ataatctgta ttagcacgaa gcatttctat tggttctttt ccaactactt
     1201 aaagcggcgc gtcgtattta atggggtatc gtttagcact tttgaggaag ccgcgctctg
     1261 cactttcttg cttaataagg agatgtattt gaaattaaga tctgacgttc tgcttccttt
     1321 aacacagtac aacagatacc tggcgttgta taacaaatac aaatatttct cgggagccat
     1381 ggatacaact tcctatagag aggctgcttg ttgccacctt gcaaaggcac tcaacgactt
     1441 cagtaacagc ggcagtgatg tgctttatca gccccctcag acttcaataa catccgccgt
     1501 cctccaagaa aacctttact ttcagtcgaa gaggcgatgg aaaaagaatt ttatcgcggt
     1561 gagtgcagcc aacagattca aaaagatatc ctcgtctggt gccctctaag ctagcttgac
     1621 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
     1681 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     1741 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     1801 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     1861 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
     1921 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
     1981 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     2041 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     2101 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
     2161 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
     2221 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
     2281 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     2341 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     2401 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2461 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2521 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2581 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2641 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2701 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2761 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2821 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2881 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2941 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3001 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3061 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3121 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3181 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3241 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     3301 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     3361 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     3421 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     3481 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg
     3541 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     3601 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     3661 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     3721 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     3781 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3841 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3901 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3961 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4021 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4081 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4141 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4201 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4261 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4321 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     4381 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     4441 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4501 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4561 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4621 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     4681 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     4741 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga
     4801 atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag
     4861 ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt
     4921 ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc
     4981 tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg
     5041 tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca
     5101 cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga
     5161 tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     5221 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     5281 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     5341 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     5401 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     5461 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga
     5521 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     5581 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     5641 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     5701 aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac
     5761 catccactcg acacacccgc cagcggccgc
//
