LOCUS       sequence-297492-        6377 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 6377)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6377)
  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-m4252820
FEATURES             Location/Qualifiers
     source          1..6377
                     /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             209..286
                     /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    339..460
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(350..369)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     404..423
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      575..621
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      651..787
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    832..903
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1129..1717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1209..1228)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2217..3426
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3263..3282
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3430..4029
                     /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(3430..3449)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3926..3946
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4056..4104
                     /note="synthetic polyadenylation signal"
     CDS             join(4370..4438,4439..5230)
                     /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(4588..4607)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5243..5379
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5531..5602
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5607..5653)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5671..5719
                     /note="synthetic polyadenylation signal"
     enhancer        5800..6103
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6105..6307
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6258..6278
                     /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 ttcagtcgaa gaggcgatgg aaaaagaatt ttatcgcggt
      241 gagtgcagcc aacagattca aaaagatatc ctcgtctggt gccctctaag ctagcttgac
      301 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
      361 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      421 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      481 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
      541 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
      601 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
      661 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
      721 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
      781 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
      841 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
      901 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
      961 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     1021 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     1081 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1141 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1201 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1261 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1321 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1381 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1441 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1501 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1561 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1621 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1681 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1741 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1801 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1861 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1921 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     1981 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     2041 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     2101 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     2161 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
     2221 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     2281 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     2341 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     2401 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     2461 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     2521 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     2581 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     2641 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     2701 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     2761 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     2821 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     2881 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     2941 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     3001 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     3061 tcacccgttc tgttggctta taatgcaggg tggggccacc agccggtagg tgtgcggtag
     3121 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     3181 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     3241 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     3301 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     3361 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     3421 ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     3481 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     3541 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     3601 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     3661 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     3721 ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg
     3781 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     3841 ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc
     3901 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     3961 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     4021 ggtgcctagg ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat
     4081 ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat
     4141 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     4201 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     4261 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     4321 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     4381 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4441 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     4501 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     4561 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4621 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     4681 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4741 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     4801 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4861 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4921 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4981 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5041 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5101 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5161 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5221 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     5281 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     5341 cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc
     5401 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     5461 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
     5521 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
     5581 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
     5641 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
     5701 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
     5761 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     5821 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     5881 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     5941 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     6001 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     6061 tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca
     6121 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     6181 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     6241 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     6301 cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca
     6361 cacccgccag cggccgc
//
