LOCUS       sequence-342387-        4590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF8 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4590)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4590)
  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-m4133532
FEATURES             Location/Qualifiers
     source          1..4590
                     /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             946..966
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             967..984
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1037..1158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1048..1067)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1102..1121
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1273..1319
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1349..1485
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1530..1601
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1827..2415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1907..1926)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3161..3229,3230..4021)
                     /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(3379..3398)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4034..4170
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4322..4393
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4398..4444)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4462..4510
                     /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 ccatgaaatt tcttgttttc
      601 ttaggaatca tcacaactgt agctgcattt caccaagaat gtagtttaca gtcatgtact
      661 caacatcaac catatgtagt tgatgacccg tgtcctattc acttctattc taaatggtat
      721 attagagtag gagctagaaa atcagcacct ttaattgaat tgtgcgtgga tgaggctggt
      781 tctaaatcac ccattcagta catcgatatc ggtaattata cagtttcctg tttacctttt
      841 acaattaatt gccaggaacc taaattgggt agtcttgtag tgcgttgttc gttctatgaa
      901 gactttttag agtatcatga cgttcgtgtt gttttagatt tcatcgaaaa cctttacttt
      961 cagtcgcacc atcaccatca ccattaagct agcttgactg actgagatac agcgtacctt
     1021 cagctcacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1081 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1141 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     1201 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct
     1261 ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt
     1321 gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac
     1381 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     1441 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc
     1501 tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac
     1561 taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg
     1621 gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt
     1681 ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt
     1741 tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac
     1801 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1861 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1921 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1981 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2041 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2101 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2161 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2221 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2281 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2341 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2401 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2461 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2521 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2581 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2641 tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc
     2701 tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg
     2761 gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa
     2821 tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc
     2881 gagatccgag gaatatcgaa atcggggcgc gcctggtgta ccgagaacga tcctctcagt
     2941 gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga
     3001 cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct
     3061 gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc
     3121 aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg
     3181 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3241 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact
     3301 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat
     3361 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3421 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac
     3481 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3541 gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac
     3601 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3661 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3721 cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac agttgataga
     3781 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3841 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3901 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3961 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4021 accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     4081 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     4141 gtgtcctcct taccagaaat ttatccttaa gatcccgaat cgtttaaact cgactctggc
     4201 tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc
     4261 gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg
     4321 acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct
     4381 tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat
     4441 gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag
     4501 cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc
     4561 attagttcat agcccatata tggagttccg
//
