LOCUS       sequence-301653-        7299 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 7299)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7299)
  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-m4252933
FEATURES             Location/Qualifiers
     source          1..7299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        97..299
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     250..270
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             557..577
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             611..688
                     /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             716..736
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1201..1322
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1212..1231)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1266..1285
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1437..1483
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1513..1649
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1694..1765
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1991..2579)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2071..2090)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3079..4288
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4125..4144
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4292..5320
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5347..5395
                     /note="synthetic polyadenylation signal"
     CDS             join(5661..5729,5730..6521)
                     /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(5879..5898)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6534..6670
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6822..6893
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6898..6944)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6962..7010
                     /note="synthetic polyadenylation signal"
     enhancer        join(7091..7299,1..95)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
       61 agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca
      121 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      181 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      241 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
      301 gtttagtgaa ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc
      361 agcggccgcc accatgtttc atctcgttga ctttcaggtt actatagcag agatattact
      421 aattattatg aggactttta aagtttccat ttggaatctt gattacatca taaacctcat
      481 aattaaaaat ttatctaagt cactaactga gaataaatat tctcaattag atgaagagca
      541 accaatggag attgatgaaa acctttactt tcagtcggga cggcgaatcc cggggctgat
      601 aaatccctgg aaaaggagat ggaagaagaa ctttattgca gtgtctgcgg ctaaccggtt
      661 caaaaagatc agttcaagcg gcgccctaga ttacgacatt cctacgaccg cctccgagaa
      721 tctctatttc cagggtgaat ttggcttggc acaacatgac gaggccgtcg ataacaagtt
      781 taataaagaa cagcagaacg ctttctatga gatccttcac ttaccaaatc tgaacgaaga
      841 gcaacgcaac gctttcattc agagcctgaa agacgatccc tcgcagtctg caaatctcct
      901 ggccgaagcc aagaagctga atgacgcaca ggctccaaaa gttgataaca agtttaacaa
      961 ggagcagcag aatgctttct acgagatatt gcaccttcct aatctgaatg aggaacaaag
     1021 aaatgcattt atccagtccc taaaagacga tccatcacag agtgccaacc tgcttgcgga
     1081 agccaaaaag ctcaatgatg cccaggcccc gaaagtagac gcgaaccatc aataaggatc
     1141 cgcaggcctc tgctagcttg actgactgag atacagcgta ccttcagctc acagacatga
     1201 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1261 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     1321 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     1381 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
     1441 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
     1501 tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
     1561 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
     1621 tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg
     1681 atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag
     1741 ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat
     1801 taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga
     1861 attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt
     1921 tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1981 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2041 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2101 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2161 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2221 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2281 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2341 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2401 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2461 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2521 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2581 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2641 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2701 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2761 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta
     2821 aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc
     2881 ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct
     2941 attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat
     3001 cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat
     3061 cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc
     3121 tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc
     3181 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     3241 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     3301 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     3361 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     3421 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     3481 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     3541 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     3601 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
     3661 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
     3721 caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc
     3781 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
     3841 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     3901 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
     3961 ccagccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
     4021 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
     4081 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
     4141 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
     4201 gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt
     4261 ctggccgttt ttggcttttt tgttagacaa catgggtaaa aagcctgaac tcaccgcgac
     4321 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     4381 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     4441 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     4501 cgccgcgctc ccgattccgg aagtgcttga cattggggag ttcagcgaga gcctgaccta
     4561 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     4621 cgctgttctg cagccggtcg cggaggcaat ggatgccatc gctgccgccg atcttagcca
     4681 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     4741 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     4801 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     4861 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     4921 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     4981 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     5041 cgagcggagg catccggagc ttgcaggatc gccccggctc cgggcgtata tgctccgcat
     5101 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     5161 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     5221 cgcccgcaga agcgccgccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     5281 aaaccgacgc cccagcactc gtccgagggc aaaggaataa gctagtatgt aagcctagtc
     5341 ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa
     5401 cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     5461 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     5521 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     5581 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     5641 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     5701 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5761 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5821 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     5881 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5941 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     6001 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6061 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6121 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6181 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     6241 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     6301 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6361 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6421 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6481 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     6541 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     6601 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     6661 ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt
     6721 tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc
     6781 tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc
     6841 cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa
     6901 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac
     6961 acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc
     7021 tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata
     7081 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     7141 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     7201 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     7261 atcatatgcc aagtacgccc cctattgacg tcaatgacg
//
