LOCUS       sequence-300449-        6278 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 6278)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6278)
  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-m4252981
FEATURES             Location/Qualifiers
     source          1..6278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(313..381,382..1173)
                     /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(531..550)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      1186..1322
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1474..1545
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1550..1596)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    1614..1662
                     /note="synthetic polyadenylation signal"
     enhancer        1743..2046
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2048..2250
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2201..2221
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2560..2681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2571..2590)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2625..2644
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2796..2842
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2872..3008
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3053..3124
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3350..3938)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3430..3449)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4438..5647
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5484..5503
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5651..6250
                     /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(5651..5670)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6147..6167
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    join(6277..6278,1..47)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 taaaatcgct atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc
       61 gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct
      121 tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt
      181 actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat
      241 cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca
      301 ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
      361 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
      421 cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
      481 agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc
      541 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
      601 cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca
      661 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
      721 ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
      781 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
      841 gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta
      901 cttactctag cttcccggca acagttgata gactggatgg aggcggataa agttgcagga
      961 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     1021 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     1081 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     1141 gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa
     1201 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
     1261 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
     1321 aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt
     1381 acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca
     1441 gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta
     1501 tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc
     1561 tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa
     1621 aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag
     1681 tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     1741 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     1801 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     1861 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     1921 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     1981 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     2041 accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     2101 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     2161 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     2221 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg
     2281 tcgatcctac catccactcg acacacccgc cagcggccgc caccatgttc cacttggtgg
     2341 attttcaggt cacgatcgca gaaatactgc taattatcat gcgaaccttc aaggttagca
     2401 tttggaacct ggactacatc attaatctca tcataaaaaa ccttagtaag tctttaactg
     2461 agaataaata ttcccaactg gatgaggaac agccaatgga gattgactaa gctagcttga
     2521 ctgactgaga tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg
     2581 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     2641 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     2701 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     2761 acaaatgtgg tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa
     2821 acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg
     2881 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag
     2941 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt
     3001 atccttaagg tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt
     3061 taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc
     3121 cctaatctca ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact
     3181 tggcgagatc gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat
     3241 ctggtccttg ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa
     3301 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3361 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3421 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3481 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3541 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3601 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3661 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3721 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3781 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3841 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3901 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3961 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4021 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4081 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4141 cagcgatctg tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc
     4201 cctcgtcgga aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg
     4261 aactatcgca agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg
     4321 ccaggtatta ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct
     4381 acatcctcaa tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc
     4441 ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca
     4501 cgtcagacga agggcgcagg agcgtcctga tccttccgcc cggacgctca ggacagcggc
     4561 ccgctgctca taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg
     4621 ggacttgggt gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa
     4681 gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt
     4741 atataaggac gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg
     4801 gttcttgttt gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc
     4861 ggggctttcg tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag
     4921 ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc
     4981 aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc
     5041 gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc aaggtcttga gcccttcgct
     5101 aatgcgggaa agctcttatt cgggtgagat gggctgggca ccatctgggg accctgacgt
     5161 gaagtttgtc actgactgga gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg
     5221 cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac
     5281 gtcacccgtt ctgttggctt ataatgcagg gtggggccac cagccggtag gtgtgcggta
     5341 ggcttttctc cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca
     5401 ggcgccggac ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt
     5461 atgtacctat cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc
     5521 gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg
     5581 taattttcag tgttagactt gtaaattgtc cgctaaattc tggccgtttt tggctttttt
     5641 gttagacaac atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     5701 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     5761 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     5821 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     5881 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     5941 gttgagcggt tcccggctgg ccgcgcagca acagatggag ggcctcctgg cgccgcaccg
     6001 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     6061 gggtctgggc agcgccgtcg tgctccccgg agtggaggcc gccgagcgcg ccggggtgcc
     6121 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     6181 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     6241 cggtgcctag gctagtatgt aagcctagtc ttagataa
//
