LOCUS       sequence-295466-        5160 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 5160)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5160)
  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-m4133960
FEATURES             Location/Qualifiers
     source          1..5160
                     /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"
     CDS             join(3153..3221,3222..4013)
                     /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(3371..3390)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4026..4162
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4314..4385
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4390..4436)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4454..4502
                     /note="synthetic polyadenylation signal"
     enhancer        4583..4886
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4888..5090
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5041..5061
                     /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 gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     2941 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     3001 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     3061 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     3121 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     3181 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3241 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     3301 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     3361 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3421 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     3481 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3541 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     3601 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3661 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     3721 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     3781 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3841 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3901 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3961 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     4021 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     4081 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     4141 cttaccagaa atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga
     4201 atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg
     4261 aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg
     4321 gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac
     4381 ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa
     4441 tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta
     4501 ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc
     4561 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     4621 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     4681 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     4741 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     4801 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4861 acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg
     4921 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     4981 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     5041 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     5101 accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
