LOCUS       sequence-297961-        4599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF7a protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4599)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4599)
  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-m4133611
FEATURES             Location/Qualifiers
     source          1..4599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    54..125
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(130..176)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    194..242
                     /note="synthetic polyadenylation signal"
     enhancer        323..626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        628..830
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     781..801
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1268..1288
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1289..1315
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1368..1489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1379..1398)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1433..1452
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1604..1650
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1680..1816
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1861..1932
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2158..2746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2238..2257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3492..3560,3561..4352)
                     /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(3710..3729)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4365..4501
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg
       61 gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac
      121 ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa
      181 tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta
      241 ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc
      301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      601 acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg
      661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
      841 accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
      901 caccatgaag atcattttgt tcctggctct tataactctc gccacctgtg agctgtacca
      961 ctatcaggaa tgcgttaggg gtacgacagt gctcttaaaa gagccttgct catcgggcac
     1021 ctatgaaggg aatagcccgt ttcatcccct ggcagataac aagtttgcgc taacatgttt
     1081 cagtactcaa tttgccttcg cttgcccaga cggagtcaaa cacgtatacc agctgagagc
     1141 acggtctgtc tcccccaagt tattcatccg ccaggaggaa gtgcaagagc tctacagccc
     1201 aatttttctt atagttgccg ctatcgtgtt cattaccctg tgttttactc tcaaacgtaa
     1261 gacagaagaa aacctttact ttcagtcgta cccctatgac gtaccagatt atgcgtaagc
     1321 tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga
     1381 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     1441 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     1501 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     1561 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg
     1621 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg
     1681 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     1741 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     1801 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga
     1861 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt
     1921 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt
     1981 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca
     2041 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat
     2101 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2161 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2221 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2281 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2341 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2401 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2461 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2521 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2581 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2641 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2701 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2761 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2821 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2881 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2941 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc
     3001 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct
     3061 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc
     3121 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag
     3181 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg
     3241 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     3301 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     3361 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     3421 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     3481 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3541 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3601 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3661 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3721 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3781 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3841 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3901 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3961 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4021 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     4081 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     4141 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4201 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4261 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4321 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     4381 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     4441 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     4501 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
     4561 cgcgaacagc cgtggcgctc atttgctcgt cgggcatcg
//
