LOCUS       sequence-300168-        6800 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 6800)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6800)
  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-m4252869
FEATURES             Location/Qualifiers
     source          1..6800
                     /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             188..265
                     /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"
     CDS             293..313
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    762..883
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(773..792)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     827..846
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      998..1044
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1074..1210
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1255..1326
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1552..2140)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1632..1651)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2640..3849
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3686..3705
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3853..4452
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3853..3872)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4349..4369
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4479..4527
                     /note="synthetic polyadenylation signal"
     CDS             join(4793..4861,4862..5653)
                     /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(5011..5030)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5666..5802
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5954..6025
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6030..6076)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6094..6142
                     /note="synthetic polyadenylation signal"
     enhancer        6223..6526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6528..6730
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6681..6701
                     /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 gtcgggacgg cgaatcccgg ggctgataaa
      181 tccctggaaa aggagatgga agaagaactt tattgcagtg tctgcggcta accggttcaa
      241 aaagatcagt tcaagcggcg ccctagatta cgacattcct acgaccgcct ccgagaatct
      301 ctatttccag ggtgaatttg gcttggcaca acatgacgag gccgtcgata acaagtttaa
      361 taaagaacag cagaacgctt tctatgagat ccttcactta ccaaatctga acgaagagca
      421 acgcaacgct ttcattcaga gcctgaaaga cgatccctcg cagtctgcaa atctcctggc
      481 cgaagccaag aagctgaatg acgcacaggc tccaaaagtt gataacaagt ttaacaagga
      541 gcagcagaat gctttctacg agatattgca ccttcctaat ctgaatgagg aacaaagaaa
      601 tgcatttatc cagtccctaa aagacgatcc atcacagagt gccaacctgc ttgcggaagc
      661 caaaaagctc aatgatgccc aggccccgaa agtagacgcg aaccatcaat aagctagctt
      721 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt
      781 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
      841 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
      901 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
      961 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct
     1021 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg
     1081 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     1141 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     1201 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc
     1261 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc
     1321 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta
     1381 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg
     1441 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc
     1501 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1561 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1621 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1681 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1741 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1801 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1861 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1921 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1981 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2041 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2101 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2161 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2221 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2281 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2341 ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag
     2401 gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa
     2461 cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat
     2521 cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac
     2581 ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg
     2641 gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc
     2701 cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct caggacagcg
     2761 gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga
     2821 cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa
     2881 aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga
     2941 ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg
     3001 cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg
     3061 ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga gagaccgcca
     3121 agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg ggggagcgca
     3181 gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg
     3241 tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt gagcccttcg
     3301 ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg ggaccctgac
     3361 gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat
     3421 ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg
     3481 acgtcacccg ttctgttggc ttataatgca gggtggggcc accagccggt aggtgtgcgg
     3541 taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga
     3601 caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt
     3661 ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg
     3721 gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata
     3781 tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt tttggctttt
     3841 ttgttagaca acatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     3901 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     3961 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     4021 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     4081 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     4141 gagttgagcg gttcccggct ggccgcgcag caacagatgg agggcctcct ggcgccgcac
     4201 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     4261 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg ccgccgagcg cgccggggtg
     4321 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     4381 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     4441 cccggtgcct aggctagtat gtaagcctag tcttagataa taaaatcgct atccatcgaa
     4501 gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc gcgcctggtg taccgagaac
     4561 gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag ccagtcggaa
     4621 tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct ggtcacggca
     4681 gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt ataatcgagt
     4741 cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc gcatgagtat
     4801 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4861 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg
     4921 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4981 ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5041 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5101 ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5161 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga ttggaggacc
     5221 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5281 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5341 aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag cttcccggca
     5401 acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5461 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5521 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5581 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5641 taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct
     5701 gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc
     5761 ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga atcgtttaaa
     5821 ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag ccgtggcgct
     5881 catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga
     5941 tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc
     6001 ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc
     6061 ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc
     6121 ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc
     6181 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     6241 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     6301 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     6361 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     6421 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     6481 tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg
     6541 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     6601 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     6661 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     6721 aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg
     6781 acacacccgc cagcggccgc
//
