LOCUS       sequence-333034-        7444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express firefly luciferase. Used in MLV-based SARS-CoV-2 pseudovirus
            assay..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7444)
  AUTHORS   Rogers TF, Zhao F, Huang D, Beutler N, Burns A, He WT, Limbo O, 
            Smith C, Song G, Woehl J, Yang L, Abbott RK, Callaghan S, Garcia E, 
            Hurtado J, Parren M, Peng L, Ramirez S, Ricketts J, Ricciardi MJ, 
            Rawlings SA, Wu NC, Yuan M, Smith DM, Nemazee D, Teijaro JR, Voss 
            JE, Wilson IA, Andrabi R, Briney B, Landais E, Sok D, Jardine JG, 
            Burton DR
  TITLE     Isolation of potent SARS-CoV-2 neutralizing antibodies and 
            protection from disease in a small animal model.
  JOURNAL   Science. 2020 Aug 21;369(6506):956-963. doi: 
            10.1126/science.abc7520. Epub 2020 Jun 15.
  PUBMED    32540903
REFERENCE   2  (bases 1 to 7444)
  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"; doi: 
            "10.1126/science.abc7520"; journalName: "Science"; date: 
            "2020-08-21- 21"; volume: "369"; issue: "6506"; pages: "956-963"
COMMENT     Sequence Label: pCMV-FLuc
FEATURES             Location/Qualifiers
     source          1..7444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    221..562
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             629..1045
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     975..997
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1013..1029
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1055..1434
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1435..1638
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1589..1609
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1635..1659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1743..1752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1749..3401
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1815..1833)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     LTR             3479..3993
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4155..4171
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4179..4185,4186..4203,4204..4209))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4224..4245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4362..4379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4533..5121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4613..4632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5292..6083,6084..6152))
                     /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     5915..5934
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6153..6257)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6325..6343
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6381..6403)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6503..6522
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6716..6738
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6866..6885
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7085..7444,1..157)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
       61 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      121 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      181 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      241 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      301 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      361 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      421 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      481 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      541 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      601 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
      661 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
      721 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
      781 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
      841 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
      901 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
      961 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1021 agccctcact ccttctctag gcgccggaat tgttgacatt gattattgac tagttattaa
     1081 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     1141 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     1201 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     1261 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     1321 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     1381 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     1441 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     1501 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     1561 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     1621 gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
     1681 tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactga gctccttaag
     1741 ccgccaccat ggaagacgcc aaaaacataa agaaaggccc ggcgccattc tatccgctgg
     1801 aagatggaac cgctggagag caactgcata aggctatgaa gagatacgcc ctggttcctg
     1861 gaacaattgc ttttacagat gcacatatcg aggtggacat cacttacgct gagtacttcg
     1921 aaatgtccgt tcggttggca gaagctatga aacgatatgg gctgaataca aatcacagaa
     1981 tcgtcgtatg cagtgaaaac tctcttcaat tctttatgcc ggtgttgggc gcgttattta
     2041 tcggagttgc agttgcgccc gcgaacgaca tttataatga acgtgaattg ctcaacagta
     2101 tgggcatttc gcagcctacc gtggtgttcg tttccaaaaa ggggttgcaa aaaattttga
     2161 acgtgcaaaa aaagctccca atcatccaaa aaattattat catggattct aaaacggatt
     2221 accagggatt tcagtcgatg tacacgttcg tcacatctca tctacctccc ggttttaatg
     2281 aatacgattt tgtgccagag tccttcgata gggacaagac aattgcactg atcatgaact
     2341 cctctggatc tactggtctg cctaaaggtg tcgctctgcc tcatagaact gcctgcgtga
     2401 gattctcgca tgccagagat cctatttttg gcaatcaaat cattccggat actgcgattt
     2461 taagtgttgt tccattccat cacggttttg gaatgtttac tacactcgga tatttgatat
     2521 gtggatttcg agtcgtctta atgtatagat ttgaagaaga gctgtttctg aggagccttc
     2581 aggattacaa gattcaaagt gcgctgctgg tgccaaccct attctccttc ttcgccaaaa
     2641 gcactctgat tgacaaatac gatttatcta atttacacga aattgcttct ggtggcgctc
     2701 ccctctctaa ggaagtcggg gaagcggttg ccaagaggtt ccatctgcca ggtatcaggc
     2761 aaggatatgg gctcactgag actacatcag ctattctgat tacacccgag ggggatgata
     2821 aaccgggcgc ggtcggtaaa gttgttccat tttttgaagc gaaggttgtg gatctggata
     2881 ccgggaaaac gctgggcgtt aatcaaagag gcgaactgtg tgtgagaggt cctatgatta
     2941 tgtccggtta tgtaaacaat ccggaagcga ccaacgcctt gattgacaag gatggatggc
     3001 tacattctgg agacatagct tactgggacg aagacgaaca cttcttcatc gttgaccgcc
     3061 tgaagtctct gattaagtac aaaggctatc aggtggctcc cgctgaattg gaatccatct
     3121 tgctccaaca ccccaacatc ttcgacgcag gtgtcgcagg tcttcccgac gatgacgccg
     3181 gtgaacttcc cgccgccgtt gttgttttgg agcacggaaa gacgatgacg gaaaaagaga
     3241 tcgtggatta cgtcgccagt caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt
     3301 ttgtggacga agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga
     3361 tcctcataaa ggccaagaag ggcggaaaga tcgccgtgta attctaggat ccgtcgacct
     3421 gcagccaagc ttatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     3481 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     3541 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag
     3601 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     3661 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
     3721 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     3781 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     3841 tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa
     3901 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     3961 gtgattgact acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa
     4021 caacattctg agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt
     4081 gaatccacat actccaatac tcctgaaata gttcattatg gacagcgcag aaagagctgg
     4141 ggagaattgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4201 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4261 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4321 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4381 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4441 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4501 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4561 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4621 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4681 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4741 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4801 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4861 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4921 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4981 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5041 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5101 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5161 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5221 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5281 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5341 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5401 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5461 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5521 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5581 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5641 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5701 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5761 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5821 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5881 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5941 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6001 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6061 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6121 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6181 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6241 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6301 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     6361 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     6421 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     6481 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     6541 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     6601 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     6661 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     6721 tcacgacgtt gtaaaacgac ggcgcaagga atggtgcatg caaggagatg gcgcccaaca
     6781 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     6841 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     6901 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggcgatta gtccaatttg
     6961 ttaaagacag gatatcagtg gtccaggctc tagttttgac tcaacaatat caccagctga
     7021 agcctataga gtacgagcca tagataaaat aaaagatttt atttagtctc cagaaaaagg
     7081 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     7141 aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag
     7201 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     7261 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
     7321 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     7381 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     7441 caac
//
