LOCUS       sequence-317276-        6341 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 6341)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6341)
  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-m4258309
FEATURES             Location/Qualifiers
     source          1..6341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             766..786
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             787..828
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    881..1002
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(892..911)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     946..965
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1117..1163
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1193..1329
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1374..1445
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1671..2259)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1751..1770)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2759..3968
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3805..3824
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3972..4571
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3972..3991)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4468..4488
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4598..4646
                     /note="synthetic polyadenylation signal"
     CDS             join(4912..4980,4981..5772)
                     /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(5130..5149)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5785..5921
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6073..6144
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6149..6195)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6213..6261
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgtttca tctcgttgac
      601 tttcaggtta ctatagcaga gatattacta attattatga ggacttttaa agtttccatt
      661 tggaatcttg attacatcat aaacctcata attaaaaatt tatctaagtc actaactgag
      721 aataaatatt ctcaattaga tgaagagcaa ccaatggaga ttgatgaaaa cctttacttt
      781 cagtcgggga aacctatacc caatccactt ctcggcttag attctaccta agctagcttg
      841 actgactgag atacagcgta ccttcagctc acagacatga taagatacat tgatgagttt
      901 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
      961 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     1021 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     1081 tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt ggggcctcta
     1141 aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac cggcattggc
     1201 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     1261 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     1321 tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca agacattcct
     1381 ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac tttcttccct
     1441 ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca agtcagctac
     1501 ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag tcaagttcga
     1561 tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat catgtgagca
     1621 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1681 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1741 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1801 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1861 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1921 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1981 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2041 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2101 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2161 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2221 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2281 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2341 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2401 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2461 tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa ctctccaagg
     2521 ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta cctctcgaac
     2581 gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc agcgcctatc
     2641 gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag aagttcaacc
     2701 tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg
     2761 cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc
     2821 acgtcagacg aagggcgcag gagcgtcctg atccttccgc ccggacgctc aggacagcgg
     2881 cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac
     2941 gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa
     3001 agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat
     3061 tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc
     3121 ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg ctgggctggc
     3181 cggggctttc gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa
     3241 gggctgtagt ctgggtccgc gagcaaggtt gccctgaact gggggttggg gggagcgcag
     3301 caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt
     3361 cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg agcccttcgc
     3421 taatgcggga aagctcttat tcgggtgaga tgggctgggc accatctggg gaccctgacg
     3481 tgaagtttgt cactgactgg agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg
     3541 gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga
     3601 cgtcacccgt tctgttggct tataatgcag ggtggggcca ccagccggta ggtgtgcggt
     3661 aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac
     3721 aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt
     3781 tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg
     3841 cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat
     3901 gtaattttca gtgttagact tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt
     3961 tgttagacaa catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     4021 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     4081 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     4141 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     4201 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     4261 agttgagcgg ttcccggctg gccgcgcagc aacagatgga gggcctcctg gcgccgcacc
     4321 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     4381 agggtctggg cagcgccgtc gtgctccccg gagtggaggc cgccgagcgc gccggggtgc
     4441 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     4501 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     4561 ccggtgccta ggctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag
     4621 atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg
     4681 atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat
     4741 ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag
     4801 cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc
     4861 ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt
     4921 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4981 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt
     5041 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc
     5101 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5161 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat
     5221 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5281 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg
     5341 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5401 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5461 atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5521 cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5581 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5641 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5701 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5761 aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg
     5821 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
     5881 cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac
     5941 tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc
     6001 atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat
     6061 cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca
     6121 taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc
     6181 caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct
     6241 tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca
     6301 attacggggt cattagttca tagcccatat atggagttcc g
//
