LOCUS       sequence-297320-        6822 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 6822)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6822)
  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-m4252937
FEATURES             Location/Qualifiers
     source          1..6822
                     /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    355..476
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(366..385)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     420..439
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      591..637
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      667..803
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    848..919
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1145..1733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1225..1244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2233..3442
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3279..3298
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3446..4474
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4501..4549
                     /note="synthetic polyadenylation signal"
     CDS             join(4815..4883,4884..5675)
                     /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(5033..5052)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5688..5824
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5976..6047
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6052..6098)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6116..6164
                     /note="synthetic polyadenylation signal"
     enhancer        6245..6548
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6550..6752
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6703..6723
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgttc cacttggtgg attttcaggt cacgatcgca gaaatactgc taattatcat
       61 gcgaaccttc aaggttagca tttggaacct ggactacatc attaatctca tcataaaaaa
      121 ccttagtaag tctttaactg agaataaata ttcccaactg gatgaggaac agccaatgga
      181 gattgacgaa aacctttact ttcagtcgaa gaggcgatgg aaaaagaatt ttatcgcggt
      241 gagtgcagcc aacagattca aaaagatatc ctcgtctggt gccctctaag gatccgcagg
      301 cctctgctag cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat
      361 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
      421 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
      481 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
      541 gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc
      601 ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat
      661 ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
      721 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
      781 ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg
      841 atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc
      901 aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca
      961 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg
     1021 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt
     1081 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1141 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1201 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1261 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1321 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1381 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1441 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1501 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1561 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1621 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1681 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1741 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1801 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     1861 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     1921 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc
     1981 cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag
     2041 gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct
     2101 tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg
     2161 agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat
     2221 cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca
     2281 cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac
     2341 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
     2401 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
     2461 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
     2521 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
     2581 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg
     2641 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt
     2701 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt
     2761 tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag
     2821 gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt
     2881 cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc
     2941 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg
     3001 gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc
     3061 tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccaccagcc
     3121 ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc
     3181 tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt
     3241 tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat
     3301 gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc
     3361 atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc
     3421 gtttttggct tttttgttag acaacatggg taaaaagcct gaactcaccg cgacgtctgt
     3481 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     3541 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     3601 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catccgccgc
     3661 gctcccgatt ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat
     3721 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     3781 tctgcagccg gtcgcggagg caatggatgc catcgctgcc gccgatctta gccagacgag
     3841 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     3901 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     3961 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     4021 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     4081 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     4141 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     4201 gaggcatccg gagcttgcag gatcgccccg gctccgggcg tatatgctcc gcattggtct
     4261 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     4321 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     4381 cagaagcgcc gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     4441 acgccccagc actcgtccga gggcaaagga ataagctagt atgtaagcct agtcttagat
     4501 aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag
     4561 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     4621 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     4681 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     4741 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     4801 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4861 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4921 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4981 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5041 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5101 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     5161 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5221 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5281 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5341 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     5401 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     5461 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5521 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5581 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5641 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     5701 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     5761 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     5821 ttaagatccc gaatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc
     5881 ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt
     5941 cagcttacct ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg
     6001 tatcttcttc cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc
     6061 cctcaagacc cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac
     6121 aaaaaaccaa cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg
     6181 agtgtagcca gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     6241 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     6301 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     6361 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     6421 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     6481 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     6541 ttaccatgct gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     6601 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     6661 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     6721 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt
     6781 tgtcgatcct accatccact cgacacaccc gccagcggcc gc
//
