LOCUS       sequence-297485-        7080 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 7080)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7080)
  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-m4252878
FEATURES             Location/Qualifiers
     source          1..7080
                     /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             503..520
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    613..734
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(624..643)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     678..697
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      849..895
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      925..1061
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1106..1177
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1403..1991)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1483..1502)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2491..3700
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3537..3556
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3704..4732
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4759..4807
                     /note="synthetic polyadenylation signal"
     CDS             join(5073..5141,5142..5933)
                     /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(5291..5310)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5946..6082
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6234..6305
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6310..6356)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6374..6422
                     /note="synthetic polyadenylation signal"
     enhancer        6503..6806
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6808..7010
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6961..6981
                     /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 gtcgatgagt gacaaaatca ttcatctaac
      181 cgatgactcc tttgatacag acgtgttgaa ggccgatggt gcgatactgg tagatttctg
      241 ggctgaatgg tgtggcccgt gcaagatgat cgcacccatt ctggacgaga tcgccgacga
      301 gtatcaagga aaattaactg tcgctaagct taatatagat cagaacccag ggacggcacc
      361 taaatacggc atcaggggaa tacctaccct cctgctcttt aagaatgggg aagttgccgc
      421 gacaaaagtg ggagccctgt ctaagggcca gctaaaagag ttcttggacg ctaacctggc
      481 aggttcaggc tcggggcaca tgcaccatca ccatcaccat agcagcgggc tcgtgccacg
      541 cggctaagga tccgcaggcc tctgctagct tgactgactg agatacagcg taccttcagc
      601 tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
      661 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
      721 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
      781 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat
      841 cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc
      901 ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg
      961 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
     1021 cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca
     1081 ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg
     1141 ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta
     1201 tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg
     1261 actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc
     1321 cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1381 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1441 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1501 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1561 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1621 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1681 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1741 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1801 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     1861 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     1921 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1981 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2041 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2101 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2161 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2221 tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc
     2281 gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct
     2341 tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc
     2401 cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga
     2461 tccgaggaat atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg gcgcctcccg
     2521 cgggcgcccc cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc aggagcgtcc
     2581 tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga
     2641 accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt
     2701 tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga
     2761 gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca
     2821 gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg
     2881 tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc
     2941 ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg
     3001 ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc cgagtcttga
     3061 atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg
     3121 gcggcaagaa cccaaggtct tgagcccttc gctaatgcgg gaaagctctt attcgggtga
     3181 gatgggctgg gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg
     3241 gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac
     3301 ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc
     3361 agggtggggc caccagccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg
     3421 ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg
     3481 ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa
     3541 gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc
     3601 accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga cttgtaaatt
     3661 gtccgctaaa ttctggccgt ttttggcttt tttgttagac aacatgggta aaaagcctga
     3721 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     3781 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     3841 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     3901 gcactttgca tccgccgcgc tcccgattcc ggaagtgctt gacattgggg agttcagcga
     3961 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     4021 aaccgaactg cccgctgttc tgcagccggt cgcggaggca atggatgcca tcgctgccgc
     4081 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     4141 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     4201 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     4261 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     4321 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     4381 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     4441 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccccggc tccgggcgta
     4501 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     4561 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     4621 gcgtacacaa atcgcccgca gaagcgccgc cgtctggacc gatggctgtg tagaagtact
     4681 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat aagctagtat
     4741 gtaagcctag tcttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt
     4801 ttgtgtgtgt aacgctaggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     4861 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     4921 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     4981 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     5041 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     5101 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5161 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     5221 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     5281 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5341 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     5401 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5461 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     5521 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5581 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     5641 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     5701 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5761 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5821 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5881 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     5941 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     6001 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     6061 cttaccagaa atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga
     6121 atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg
     6181 aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg
     6241 gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac
     6301 ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa
     6361 tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta
     6421 ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc
     6481 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     6541 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     6601 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     6661 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     6721 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     6781 acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg
     6841 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     6901 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     6961 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     7021 accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
