LOCUS       sequence-297497-        6768 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 6768)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6768)
  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-m4252880
FEATURES             Location/Qualifiers
     source          1..6768
                     /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..232
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    301..422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(312..331)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     366..385
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      537..583
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      613..749
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    794..865
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1091..1679)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1171..1190)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2179..3388
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3225..3244
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3392..4420
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4447..4495
                     /note="synthetic polyadenylation signal"
     CDS             join(4761..4829,4830..5621)
                     /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(4979..4998)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5634..5770
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5922..5993
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5998..6044)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6062..6110
                     /note="synthetic polyadenylation signal"
     enhancer        6191..6494
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6496..6698
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6649..6669
                     /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 gtcgaagagg cgatggaaaa agaattttat
      181 cgcggtgagt gcagccaaca gattcaaaaa gatatcctcg tctggtgccc tctaaggatc
      241 cgcaggcctc tgctagcttg actgactgag atacagcgta ccttcagctc acagacatga
      301 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      361 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      421 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
      481 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
      541 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
      601 tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
      661 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
      721 tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg
      781 atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag
      841 ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat
      901 taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga
      961 attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt
     1021 tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1081 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1141 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1201 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1261 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1321 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1381 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1441 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1501 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     1561 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1621 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1681 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1741 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1801 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     1861 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta
     1921 aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc
     1981 ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct
     2041 attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat
     2101 cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat
     2161 cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc
     2221 tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc
     2281 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     2341 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     2401 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     2461 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     2521 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     2581 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     2641 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     2701 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
     2761 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
     2821 caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc
     2881 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
     2941 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     3001 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
     3061 ccagccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
     3121 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
     3181 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
     3241 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
     3301 gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt
     3361 ctggccgttt ttggcttttt tgttagacaa catgggtaaa aagcctgaac tcaccgcgac
     3421 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     3481 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     3541 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     3601 cgccgcgctc ccgattccgg aagtgcttga cattggggag ttcagcgaga gcctgaccta
     3661 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     3721 cgctgttctg cagccggtcg cggaggcaat ggatgccatc gctgccgccg atcttagcca
     3781 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     3841 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     3901 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     3961 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     4021 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     4081 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     4141 cgagcggagg catccggagc ttgcaggatc gccccggctc cgggcgtata tgctccgcat
     4201 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     4261 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     4321 cgcccgcaga agcgccgccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     4381 aaaccgacgc cccagcactc gtccgagggc aaaggaataa gctagtatgt aagcctagtc
     4441 ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa
     4501 cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     4561 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     4621 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     4681 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     4741 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     4801 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4861 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4921 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     4981 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5041 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     5101 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5161 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5221 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5281 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     5341 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     5401 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5461 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5521 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5581 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     5641 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     5701 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     5761 ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt
     5821 tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc
     5881 tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc
     5941 cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa
     6001 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac
     6061 acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc
     6121 tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata
     6181 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     6241 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     6301 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     6361 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     6421 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     6481 tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     6541 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     6601 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     6661 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca
     6721 gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgc
//
