LOCUS       sequence-295188-        4947 bp    DNA     circular SYN 15-DEC-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 4947)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4947)
  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-m4133840
FEATURES             Location/Qualifiers
     source          1..4947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             188..208
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             242..319
                     /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"
     CDS             347..367
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    816..937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(827..846)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     881..900
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1052..1098
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1128..1264
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1309..1380
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1606..2194)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1686..1705)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2940..3008,3009..3800)
                     /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(3158..3177)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3813..3949
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4101..4172
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4177..4223)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4241..4289
                     /note="synthetic polyadenylation signal"
     enhancer        4370..4673
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4675..4877
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4828..4848
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgttt catctcgttg actttcaggt tactatagca gagatattac taattattat
       61 gaggactttt aaagtttcca tttggaatct tgattacatc ataaacctca taattaaaaa
      121 tttatctaag tcactaactg agaataaata ttctcaatta gatgaagagc aaccaatgga
      181 gattgatgaa aacctttact ttcagtcggg acggcgaatc ccggggctga taaatccctg
      241 gaaaaggaga tggaagaaga actttattgc agtgtctgcg gctaaccggt tcaaaaagat
      301 cagttcaagc ggcgccctag attacgacat tcctacgacc gcctccgaga atctctattt
      361 ccagggtgaa tttggcttgg cacaacatga cgaggccgtc gataacaagt ttaataaaga
      421 acagcagaac gctttctatg agatccttca cttaccaaat ctgaacgaag agcaacgcaa
      481 cgctttcatt cagagcctga aagacgatcc ctcgcagtct gcaaatctcc tggccgaagc
      541 caagaagctg aatgacgcac aggctccaaa agttgataac aagtttaaca aggagcagca
      601 gaatgctttc tacgagatat tgcaccttcc taatctgaat gaggaacaaa gaaatgcatt
      661 tatccagtcc ctaaaagacg atccatcaca gagtgccaac ctgcttgcgg aagccaaaaa
      721 gctcaatgat gcccaggccc cgaaagtaga cgcgaaccat caataagcta gcttgactga
      781 ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca
      841 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
      901 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
      961 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     1021 atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg
     1081 gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga
     1141 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     1201 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     1261 ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat
     1321 ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta
     1381 atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc
     1441 gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg
     1501 tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg
     1561 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1621 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1681 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1741 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1801 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1861 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1921 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1981 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2041 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2101 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2161 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2221 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2281 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2341 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2401 gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc
     2461 gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact
     2521 atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag
     2581 gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat
     2641 cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggtgtac
     2701 cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca
     2761 gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt
     2821 cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata
     2881 atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca
     2941 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3001 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc
     3061 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3121 aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3181 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3241 ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3301 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg
     3361 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3421 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3481 ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt
     3541 cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3601 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3661 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3721 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3781 cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg
     3841 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     3901 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc
     3961 gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg
     4021 tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg
     4081 gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct tcttccctct
     4141 agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt
     4201 agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac
     4261 atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct
     4321 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     4381 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     4441 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     4501 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     4561 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     4621 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc
     4681 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     4741 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     4801 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     4861 tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat
     4921 ccactcgaca cacccgccag cggccgc
//
