LOCUS       sequence-295038-        6847 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 6847)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6847)
  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-m4133239
FEATURES             Location/Qualifiers
     source          1..6847
                     /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"
     promoter        2694..3903
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3740..3759
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3907..4506
                     /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(3907..3926)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4403..4423
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4533..4581
                     /note="synthetic polyadenylation signal"
     CDS             join(4847..4915,4916..5707)
                     /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(5065..5084)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5720..5856
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6001..6072
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6077..6123)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6141..6189
                     /note="synthetic polyadenylation signal"
     enhancer        6270..6573
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6575..6777
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6728..6748
                     /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 cctggcctcc
     2701 gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc
     2761 agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc
     2821 tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac
     2881 ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag
     2941 tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat
     3001 aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc
     3061 ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg
     3121 ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct
     3181 gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa
     3241 tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg
     3301 aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg
     3361 cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc tgacgtgaag
     3421 tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt
     3481 gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca
     3541 cccgttctgt tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct
     3601 tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg
     3661 ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt
     3721 acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt
     3781 gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat
     3841 tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc ttttttgtta
     3901 gacaacatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     3961 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     4021 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     4081 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     4141 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     4201 agcggttccc ggctggccgc gcagcaacag atggagggcc tcctggcgcc gcaccggccc
     4261 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     4321 ctgggcagcg ccgtcgtgct ccccggagtg gaggccgccg agcgcgccgg ggtgcccgcc
     4381 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     4441 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     4501 gcctaggcta gtatgtaagc ctagtcttag ataataaaat cgctatccat cgaagatgga
     4561 tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct
     4621 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     4681 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     4741 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     4801 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca
     4861 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4921 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat
     4981 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc
     5041 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5101 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc
     5161 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5221 aaccatgagt gataacactg cggccaactt acttctgaca acgattggag gaccgaagga
     5281 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5341 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5401 aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt
     5461 gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5521 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5581 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5641 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5701 ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     5761 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     5821 ccatacgtgt cctccttacc agaaatttat ccttaagatc gtttaaactc gactctggct
     5881 ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg
     5941 ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga
     6001 catcttggac cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt
     6061 cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg
     6121 ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc
     6181 gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca
     6241 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     6301 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     6361 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     6421 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     6481 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     6541 tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat
     6601 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     6661 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     6721 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
     6781 ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag
     6841 cggccgc
//
