LOCUS       sequence-297496-        6708 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 6708)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6708)
  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-m4252957
FEATURES             Location/Qualifiers
     source          1..6708
                     /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             155..172
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    241..362
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(252..271)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     306..325
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      477..523
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      553..689
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    734..805
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1031..1619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1111..1130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2119..3328
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3165..3184
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3332..4360
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4387..4435
                     /note="synthetic polyadenylation signal"
     CDS             join(4701..4769,4770..5561)
                     /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(4919..4938)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5574..5710
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5862..5933
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5938..5984)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6002..6050
                     /note="synthetic polyadenylation signal"
     enhancer        6131..6434
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6436..6638
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6589..6609
                     /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 gccgaaaacc tttactttca gtcgcaccat caccatcacc attaaggatc
      181 cgcaggcctc tgctagcttg actgactgag atacagcgta ccttcagctc acagacatga
      241 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      301 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      361 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
      421 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
      481 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
      541 tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
      601 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
      661 tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg
      721 atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag
      781 ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat
      841 taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga
      901 attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt
      961 tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1021 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1081 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1141 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1201 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1261 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1321 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1381 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1441 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     1501 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1561 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1621 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1681 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1741 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     1801 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta
     1861 aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc
     1921 ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct
     1981 attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat
     2041 cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat
     2101 cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc
     2161 tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc
     2221 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     2281 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     2341 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     2401 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     2461 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     2521 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     2581 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     2641 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
     2701 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
     2761 caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc
     2821 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
     2881 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     2941 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
     3001 ccagccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
     3061 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
     3121 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
     3181 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
     3241 gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt
     3301 ctggccgttt ttggcttttt tgttagacaa catgggtaaa aagcctgaac tcaccgcgac
     3361 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     3421 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     3481 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     3541 cgccgcgctc ccgattccgg aagtgcttga cattggggag ttcagcgaga gcctgaccta
     3601 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     3661 cgctgttctg cagccggtcg cggaggcaat ggatgccatc gctgccgccg atcttagcca
     3721 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     3781 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     3841 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     3901 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     3961 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     4021 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     4081 cgagcggagg catccggagc ttgcaggatc gccccggctc cgggcgtata tgctccgcat
     4141 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     4201 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     4261 cgcccgcaga agcgccgccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     4321 aaaccgacgc cccagcactc gtccgagggc aaaggaataa gctagtatgt aagcctagtc
     4381 ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa
     4441 cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     4501 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     4561 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     4621 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     4681 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     4741 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4801 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4861 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     4921 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4981 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     5041 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5101 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5161 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5221 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     5281 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     5341 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5401 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5461 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5521 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     5581 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     5641 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     5701 ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt
     5761 tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc
     5821 tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc
     5881 cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa
     5941 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac
     6001 acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc
     6061 tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata
     6121 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     6181 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     6241 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     6301 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     6361 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     6421 tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     6481 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     6541 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     6601 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca
     6661 gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgc
//
