LOCUS       sequence-342382-        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-m4133249
FEATURES             Location/Qualifiers
     source          1..4590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        76..379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     534..554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1021..1041
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1042..1059
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1112..1233
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1123..1142)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1177..1196
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1348..1394
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1424..1560
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1605..1676
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1902..2490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1982..2001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3236..3304,3305..4096)
                     /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(3454..3473)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4109..4245
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4397..4468
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4473..4519)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4537..4585
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc
       61 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      121 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      181 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      241 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      301 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      361 agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      421 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      481 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      541 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      601 gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgccaccatg
      661 aaattcttag tgtttcttgg gatcattaca acggtcgcag cttttcatca ggaatgcagc
      721 ctacaatcgt gtacccagca ccagccgtac gttgtggacg atccttgccc catccacttc
      781 tattctaagt ggtacatacg ggtaggcgcg aggaagagtg caccactcat tgagttgtgt
      841 gtggacgagg ccggttcaaa atccccaatc cagtatatag atattggaaa ttacactgtc
      901 tcctgtctgc ccttcaccat caactgccaa gaacctaagc tgggaagcct cgttgtgaga
      961 tgctcttttt atgaggattt tcttgaatac cacgacgtcc gtgtggttct ggacttcatt
     1021 gaaaaccttt actttcagtc gcaccatcac catcaccatt aagctagctt gactgactga
     1081 gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt tggacaaacc
     1141 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     1201 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     1261 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     1321 ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct aaacgggtct
     1381 tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa
     1441 aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat
     1501 acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa
     1561 ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc
     1621 ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct
     1681 cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga
     1741 tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg atctggtcct
     1801 tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc aaaaggccag
     1861 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1921 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1981 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2041 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2101 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2161 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2221 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2281 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2341 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2401 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2461 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2521 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2581 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2641 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2701 tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag gccctcgtcg
     2761 gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa cgaactatcg
     2821 caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat cgccaggtat
     2881 tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac ctacatcctc
     2941 aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg gtgtaccgag
     3001 aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg
     3061 gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg
     3121 gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg
     3181 agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag
     3241 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3301 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt
     3361 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3421 acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3481 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3541 gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3601 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg
     3661 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3721 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3781 agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg
     3841 gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3901 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3961 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4021 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4081 gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac
     4141 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     4201 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt
     4261 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     4321 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     4381 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     4441 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     4501 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     4561 atcttcgatg gatagcgatt ttattatcta
//
