LOCUS       sequence-295174-        4782 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 4782)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4782)
  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-m4133431
FEATURES             Location/Qualifiers
     source          1..4782
                     /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             557..574
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    651..772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(662..681)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     716..735
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      887..933
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      963..1099
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1144..1215
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1441..2029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1521..1540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2775..2843,2844..3635)
                     /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(2993..3012)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3648..3784
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3936..4007
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4012..4058)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4076..4124
                     /note="synthetic polyadenylation signal"
     enhancer        4205..4508
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4510..4712
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4663..4683
                     /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 ttcagtcgat gagtgacaaa atcattcatc taaccgatga
      241 ctcctttgat acagacgtgt tgaaggccga tggtgcgata ctggtagatt tctgggctga
      301 atggtgtggc ccgtgcaaga tgatcgcacc cattctggac gagatcgccg acgagtatca
      361 aggaaaatta actgtcgcta agcttaatat agatcagaac ccagggacgg cacctaaata
      421 cggcatcagg ggaataccta ccctcctgct ctttaagaat ggggaagttg ccgcgacaaa
      481 agtgggagcc ctgtctaagg gccagctaaa agagttcttg gacgctaacc tggcaggttc
      541 aggctcgggg cacatgcacc atcaccatca ccatagcagc gggctcgtgc cacgcggcta
      601 agctagcttg actgactgag atacagcgta ccttcagctc acagacatga taagatacat
      661 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
      721 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
      781 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
      841 gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt
      901 ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac
      961 cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     1021 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     1081 accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca
     1141 agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac
     1201 tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca
     1261 agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag
     1321 tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat
     1381 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1441 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1501 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1561 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1621 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1681 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1741 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1801 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1861 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     1921 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1981 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2041 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2101 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2161 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2221 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa
     2281 ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta
     2341 cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc
     2401 agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag
     2461 aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg
     2521 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     2581 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     2641 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     2701 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     2761 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2821 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2881 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2941 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3001 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3061 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     3121 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3181 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3241 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3301 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     3361 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     3421 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3481 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3541 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3601 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     3661 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     3721 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     3781 ttaagatccc gaatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc
     3841 ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt
     3901 cagcttacct ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg
     3961 tatcttcttc cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc
     4021 cctcaagacc cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac
     4081 aaaaaaccaa cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg
     4141 agtgtagcca gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     4201 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     4261 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     4321 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     4381 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     4441 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     4501 ttaccatgct gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     4561 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     4621 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     4681 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt
     4741 tgtcgatcct accatccact cgacacaccc gccagcggcc gc
//
