LOCUS       sequence-297403-        6813 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 6813)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6813)
  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-m4252925
FEATURES             Location/Qualifiers
     source          1..6813
                     /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             212..277
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    346..467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(357..376)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     411..430
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      582..628
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      658..794
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    839..910
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1136..1724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1216..1235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2224..3433
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3270..3289
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3437..4465
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4492..4540
                     /note="synthetic polyadenylation signal"
     CDS             join(4806..4874,4875..5666)
                     /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(5024..5043)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5679..5815
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5967..6038
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6043..6089)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6107..6155
                     /note="synthetic polyadenylation signal"
     enhancer        6236..6539
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6541..6743
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6694..6714
                     /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 ttcagtcgat ggattataag gaccacgatg gagactacaa
      241 agatcatgac attgattaca aggacgacga tgacaaataa ggatccgcag gcctctgcta
      301 gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg
      361 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
      421 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
      481 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
      541 acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc
      601 ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca
      661 ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag
      721 attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag
      781 aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca
      841 ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct
      901 tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca
      961 gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag
     1021 ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt
     1081 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1141 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1201 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1261 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1321 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1381 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1441 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1501 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1561 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1621 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1681 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1741 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1801 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     1861 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1921 ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc
     1981 caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct
     2041 cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc
     2101 ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt
     2161 caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg
     2221 cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg
     2281 ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac
     2341 agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt
     2401 aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga
     2461 ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg
     2521 atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg
     2581 gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg
     2641 ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc
     2701 gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag
     2761 cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt
     2821 gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc
     2881 ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc
     2941 tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag
     3001 ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt
     3061 cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccaccagc cggtaggtgt
     3121 gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa
     3181 tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc
     3241 ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg
     3301 gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc
     3361 aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc
     3421 ttttttgtta gacaacatgg gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt
     3481 tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc
     3541 tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc
     3601 cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatccgccg cgctcccgat
     3661 tccggaagtg cttgacattg gggagttcag cgagagcctg acctattgca tctcccgccg
     3721 tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc
     3781 ggtcgcggag gcaatggatg ccatcgctgc cgccgatctt agccagacga gcgggttcgg
     3841 cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat
     3901 tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt
     3961 cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct
     4021 cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt
     4081 cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt
     4141 ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc
     4201 ggagcttgca ggatcgcccc ggctccgggc gtatatgctc cgcattggtc ttgaccaact
     4261 ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga
     4321 cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc
     4381 cgccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag
     4441 cactcgtccg agggcaaagg aataagctag tatgtaagcc tagtcttaga taataaaatc
     4501 gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg
     4561 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt
     4621 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag
     4681 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa
     4741 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct
     4801 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4861 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4921 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4981 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5041 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5101 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5161 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5221 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5281 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5341 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc
     5401 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc
     5461 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5521 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5581 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5641 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc
     5701 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     5761 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc
     5821 cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa
     5881 cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc
     5941 tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt
     6001 ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac
     6061 ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca
     6121 acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc
     6181 agatctagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     6241 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     6301 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     6361 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     6421 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     6481 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
     6541 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     6601 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     6661 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     6721 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc
     6781 taccatccac tcgacacacc cgccagcggc cgc
//
