LOCUS       sequence-300596-        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"
     polyA_signal    35..156
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(46..65)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     100..119
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      271..317
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      347..483
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    528..599
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(825..1413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(905..924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1913..3122
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2959..2978
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3126..4154
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4181..4229
                     /note="synthetic polyadenylation signal"
     CDS             join(4495..4563,4564..5355)
                     /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(4713..4732)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5368..5504
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5656..5727
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5732..5778)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5796..5844
                     /note="synthetic polyadenylation signal"
     enhancer        5925..6228
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6230..6432
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6383..6403
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             6690..6710
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             6711..6788
                     /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"
ORIGIN
        1 tgagatacag cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa
       61 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
      121 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
      181 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
      241 tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg
      301 tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa
      361 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
      421 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
      481 taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg
      541 gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa
      601 tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg
      661 agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt
      721 ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc
      781 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
      841 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
      901 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
      961 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1021 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1081 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1141 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     1201 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     1261 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     1321 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     1381 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     1441 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     1501 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     1561 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     1621 atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg
     1681 tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta
     1741 tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg
     1801 tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc
     1861 ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg
     1921 cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca
     1981 gacgaagggc gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct
     2041 gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact
     2101 tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt
     2161 cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata
     2221 aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct
     2281 tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc
     2341 tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg
     2401 tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat
     2461 ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga
     2521 aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc
     2581 gggaaagctc ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt
     2641 ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg
     2701 ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac
     2761 ccgttctgtt ggcttataat gcagggtggg gccaccagcc ggtaggtgtg cggtaggctt
     2821 ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc
     2881 cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta
     2941 cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg
     3001 tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt
     3061 ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag
     3121 acaacatggg taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
     3181 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
     3241 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
     3301 acaaagatcg ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc
     3361 ttgacattgg ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
     3421 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
     3481 caatggatgc catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac
     3541 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
     3601 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
     3661 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     3721 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     3781 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     3841 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     3901 gatcgccccg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     3961 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     4021 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga
     4081 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     4141 gggcaaagga ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg
     4201 aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga
     4261 acgatcctct cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg
     4321 aatccagctt gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg
     4381 cagcgtaccg atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga
     4441 gtcctagctt ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt
     4501 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4561 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg
     4621 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4681 cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4741 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4801 tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4861 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga
     4921 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4981 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5041 gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg
     5101 caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5161 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5221 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5281 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5341 attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg
     5401 ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct tccccaactt
     5461 gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta
     5521 aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg
     5581 ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc
     5641 gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg
     5701 tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg
     5761 ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca
     5821 tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa
     5881 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     5941 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     6001 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     6061 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     6121 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     6181 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt
     6241 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     6301 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     6361 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     6421 ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact
     6481 cgacacaccc gccagcggcc gccaccatgt tccacttggt ggattttcag gtcacgatcg
     6541 cagaaatact gctaattatc atgcgaacct tcaaggttag catttggaac ctggactaca
     6601 tcattaatct catcataaaa aaccttagta agtctttaac tgagaataaa tattcccaac
     6661 tggatgagga acagccaatg gagattgacg aaaaccttta ctttcagtcg aagaggcgat
     6721 ggaaaaagaa ttttatcgcg gtgagtgcag ccaacagatt caaaaagata tcctcgtctg
     6781 gtgccctcta aggatccgca ggcctctgct agcttgactg ac
//
