LOCUS       sequence-297328-        6759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF7b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6759)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6759)
  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-m4252967
FEATURES             Location/Qualifiers
     source          1..6759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             134..154
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             158..223
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    292..413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(303..322)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     357..376
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      528..574
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      604..740
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    785..856
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1082..1670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1162..1181)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2170..3379
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3216..3235
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3383..4411
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4438..4486
                     /note="synthetic polyadenylation signal"
     CDS             join(4752..4820,4821..5612)
                     /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(4970..4989)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5625..5761
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5913..5984
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5989..6035)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6053..6101
                     /note="synthetic polyadenylation signal"
     enhancer        6182..6485
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6487..6689
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6640..6660
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgata gaactatcgc tcatcgactt ttatctgtgc ttcttggctt tcctgctttt
       61 tctggtgctc attatgctaa tcattttttg gttcagtctg gagttacaag atcataatga
      121 gacatgtcac gccgaaaacc tttactttca gtcgatggat tataaggacc acgatggaga
      181 ctacaaagat catgacattg attacaagga cgacgatgac aaataaggat ccgcaggcct
      241 ctgctagctt gactgactga gatacagcgt accttcagct cacagacatg ataagataca
      301 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
      361 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
      421 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
      481 agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct
      541 tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta
      601 ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
      661 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
      721 taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc
      781 aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa
      841 ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc
      901 aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca
      961 gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa
     1021 tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1081 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1141 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1201 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1261 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1321 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1381 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1441 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1501 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1561 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1621 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1681 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1741 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1801 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1861 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga
     1921 actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct
     1981 acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg
     2041 cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga
     2101 gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg
     2161 ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg
     2221 cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct
     2281 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga
     2341 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac
     2401 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga
     2461 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg
     2521 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct
     2581 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga
     2641 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg
     2701 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg
     2761 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt
     2821 gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg
     2881 ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg
     2941 cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg
     3001 tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc accagccggt
     3061 aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct
     3121 cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct
     3181 ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg
     3241 ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt
     3301 tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt
     3361 tttggctttt ttgttagaca acatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga
     3421 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     3481 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     3541 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat ccgccgcgct
     3601 cccgattccg gaagtgcttg acattgggga gttcagcgag agcctgacct attgcatctc
     3661 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     3721 gcagccggtc gcggaggcaa tggatgccat cgctgccgcc gatcttagcc agacgagcgg
     3781 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     3841 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     3901 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     3961 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
     4021 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
     4081 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
     4141 gcatccggag cttgcaggat cgccccggct ccgggcgtat atgctccgca ttggtcttga
     4201 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
     4261 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
     4321 aagcgccgcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
     4381 ccccagcact cgtccgaggg caaaggaata agctagtatg taagcctagt cttagataat
     4441 aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg
     4501 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     4561 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     4621 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     4681 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     4741 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4801 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4861 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4921 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4981 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5041 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5101 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5161 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5221 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5281 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     5341 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     5401 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5461 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5521 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5581 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     5641 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     5701 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     5761 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
     5821 cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag
     5881 cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat
     5941 cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct
     6001 caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa
     6061 aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt
     6121 gtagccagat ctagtaatca attacggggt cattagttca tagcccatat atggagttcc
     6181 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     6241 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     6301 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     6361 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     6421 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     6481 ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     6541 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     6601 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     6661 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt
     6721 cgatcctacc atccactcga cacacccgcc agcggccgc
//
