LOCUS       sequence-297540-        6669 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 6669)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6669)
  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-m4252985
FEATURES             Location/Qualifiers
     source          1..6669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    202..323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(213..232)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     267..286
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      438..484
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      514..650
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    695..766
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(992..1580)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1072..1091)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2080..3289
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3126..3145
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3293..4321
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4348..4396
                     /note="synthetic polyadenylation signal"
     CDS             join(4662..4730,4731..5522)
                     /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(4880..4899)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5535..5671
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5823..5894
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5899..5945)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5963..6011
                     /note="synthetic polyadenylation signal"
     enhancer        6092..6395
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6397..6599
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6550..6570
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgatt gaactttcat taattgactt ctatttgtgc tttttagcct ttctgctatt
       61 ccttgtttta attatgctta ttatcttttg gttctcactt gaactgcaag atcataatga
      121 aacttgtcac gcctaaggat ccgcaggcct ctgctagctt gactgactga gatacagcgt
      181 accttcagct cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
      241 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
      301 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
      361 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc
      421 catctctatc ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt
      481 tttgtgcccc tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat
      541 gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc
      601 caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc
      661 tatcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac
      721 accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac
      781 cttgggctat cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc
      841 tgggtttcga ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca
      901 cccgttctcc gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca
      961 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1021 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1081 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1141 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1201 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1261 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1321 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1381 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     1441 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1501 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1561 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1621 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1681 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1741 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     1801 gttcatccat agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc
     1861 aaacctttcg tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt
     1921 ggccggcctt gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc
     1981 ccgaatatcc gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc
     2041 tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg
     2101 cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca
     2161 ggagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc
     2221 ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag
     2281 ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga
     2341 ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg
     2401 tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg
     2461 ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc
     2521 gggccgctcg gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg
     2581 cgagcaaggt tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc
     2641 gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg
     2701 gcatggtggg cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg aaagctctta
     2761 ttcgggtgag atgggctggg caccatctgg ggaccctgac gtgaagtttg tcactgactg
     2821 gagaactcgg tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg
     2881 cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc
     2941 ttataatgca gggtggggcc accagccggt aggtgtgcgg taggcttttc tccgtcgcag
     3001 gacgcagggt tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg
     3061 aggggaggga taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa
     3121 gtagctgaag ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt
     3181 tttttaggca ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac
     3241 ttgtaaattg tccgctaaat tctggccgtt tttggctttt ttgttagaca acatgggtaa
     3301 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     3361 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     3421 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     3481 tgtttatcgg cactttgcat ccgccgcgct cccgattccg gaagtgcttg acattgggga
     3541 gttcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     3601 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcaa tggatgccat
     3661 cgctgccgcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     3721 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     3781 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     3841 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     3901 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     3961 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     4021 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccccggct
     4081 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     4141 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     4201 gactgtcggg cgtacacaaa tcgcccgcag aagcgccgcc gtctggaccg atggctgtgt
     4261 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     4321 agctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt
     4381 gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag
     4441 tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg
     4501 acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc
     4561 tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg
     4621 caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc
     4681 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4741 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac
     4801 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga
     4861 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4921 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca
     4981 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5041 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa
     5101 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5161 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5221 ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag
     5281 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5341 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5401 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5461 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5521 aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     5581 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     5641 cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg
     5701 ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt
     5761 cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc
     5821 gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc
     5881 ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta
     5941 tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata
     6001 gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt
     6061 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     6121 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     6181 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     6241 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     6301 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     6361 agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca
     6421 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     6481 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     6541 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     6601 gtttagtgaa ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc
     6661 agcggccgc
//
