LOCUS       sequence-317729-        6969 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 6969)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6969)
  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-m4133801
FEATURES             Location/Qualifiers
     source          1..6969
                     /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..1011
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1080..1201
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1091..1110)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1145..1164
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1316..1362
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1392..1528
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1573..1644
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1870..2458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1950..1969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2958..4167
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4004..4023
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4171..5199
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5226..5274
                     /note="synthetic polyadenylation signal"
     CDS             join(5540..5608,5609..6400)
                     /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(5758..5777)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6413..6549
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6701..6772
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6777..6823)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6841..6889
                     /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 ccatgaagat cattttgttc
      601 ctggctctta taactctcgc cacctgtgag ctgtaccact atcaggaatg cgttaggggt
      661 acgacagtgc tcttaaaaga gccttgctca tcgggcacct atgaagggaa tagcccgttt
      721 catcccctgg cagataacaa gtttgcgcta acatgtttca gtactcaatt tgccttcgct
      781 tgcccagacg gagtcaaaca cgtataccag ctgagagcac ggtctgtctc ccccaagtta
      841 ttcatccgcc aggaggaagt gcaagagctc tacagcccaa tttttcttat agttgccgct
      901 atcgtgttca ttaccctgtg ttttactctc aaacgtaaga cagaagaaaa cctttacttt
      961 cagtcgaaag aaaccgcggc agctaagttc gagaggcagc acatggacag ttaaggatcc
     1021 gcaggcctct gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat
     1081 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1141 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1201 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1261 ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca
     1321 taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt
     1381 gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     1441 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     1501 gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga
     1561 tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt
     1621 tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt
     1681 aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa
     1741 ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt
     1801 catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1861 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1921 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1981 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2041 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2101 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2161 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2221 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2281 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2341 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2401 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2461 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2521 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2581 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2641 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa
     2701 atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct
     2761 tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta
     2821 ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc
     2881 acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc
     2941 gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
     3001 cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga tccttccgcc
     3061 cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag
     3121 cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag
     3181 agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg
     3241 ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg
     3301 cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt
     3361 agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag
     3421 cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg
     3481 ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt
     3541 gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc
     3601 aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat gggctgggca
     3661 ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt
     3721 tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg
     3781 cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac
     3841 ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg
     3901 taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt
     3961 cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga
     4021 actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg
     4081 taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc cgctaaattc
     4141 tggccgtttt tggctttttt gttagacaac atgggtaaaa agcctgaact caccgcgacg
     4201 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
     4261 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
     4321 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcc
     4381 gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag cctgacctat
     4441 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
     4501 gctgttctgc agccggtcgc ggaggcaatg gatgccatcg ctgccgccga tcttagccag
     4561 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
     4621 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     4681 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     4741 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
     4801 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
     4861 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     4921 gagcggaggc atccggagct tgcaggatcg ccccggctcc gggcgtatat gctccgcatt
     4981 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     5041 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     5101 gcccgcagaa gcgccgccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     5161 aaccgacgcc ccagcactcg tccgagggca aaggaataag ctagtatgta agcctagtct
     5221 tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac
     5281 gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     5341 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     5401 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     5461 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     5521 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     5581 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5641 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5701 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     5761 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5821 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     5881 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5941 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6001 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6061 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     6121 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     6181 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6241 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6301 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6361 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     6421 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     6481 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     6541 tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     6601 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     6661 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     6721 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     6781 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     6841 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     6901 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     6961 ggagttccg
//
