LOCUS       sequence-348112-        7722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient mammalian expression of Luc2 (firefly luciferase). Lacks 
            packaging sequence and is used as a negative control transfer 
            plasmid for SARS-CoV-2 virus-like particles..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7722)
  AUTHORS   Syed AM, Taha TY, Tabata T, Chen IP, Ciling A, Khalid MM, Sreekumar 
            B, Chen PY, Hayashi JM, Soczek KM, Ott M, Doudna JA
  TITLE     Rapid assessment of SARS-CoV-2 evolved variants using virus-like 
            particles.
  JOURNAL   Science. 2021 Nov 4:eabl6184. doi: 10.1126/science.abl6184.
  PUBMED    34735219
REFERENCE   2  (bases 1 to 7722)
  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"; journalName: "Science. 
            2021 Nov 4:eabl6184. doi: 10.1126/science.abl6184."
COMMENT     Sequence Label: Luc-noPS
FEATURES             Location/Qualifiers
     source          1..7722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        129..508
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        509..712
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     663..683
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     757..776
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        757..775
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      795..804
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             801..2453
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     misc_feature    2550..3138
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2603..2623)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3021..3032)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3207..3224)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3213..3437
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3483..3911
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3570..3589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3780..3801
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3920..3940)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3925..4254
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4105..4240
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4167..4186
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4321..5115
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4375..4394)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4985..5004
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5289..5410
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5326..5345)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5380..5399
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5459..5475)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5459..5475)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5472..5494)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5483..5499
                     /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(5507..5513,5514..5531,5532..5537))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5552..5573
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5690..5707)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5861..6449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5941..5960)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6620..7411,7412..7480))
                     /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     7243..7262
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7481..7585)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7660..7679)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
       61 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
      121 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
      181 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      241 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      301 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      361 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      421 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      481 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      541 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      601 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      661 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct
      721 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
      781 ccaagctggc tagcgccgcc atggaagatg ccaaaaacat taagaagggc ccagcgccat
      841 tctacccact cgaagacggg accgccggcg agcagctgca caaagccatg aagcgctacg
      901 ccctggtgcc cggcaccatc gcctttaccg acgcacatat cgaggtggac attacctacg
      961 ccgagtactt cgagatgagc gttcggctgg cagaagctat gaagcgctat gggctgaata
     1021 caaaccatcg gatcgtggtg tgcagcgaga atagcttgca gttcttcatg cccgtgttgg
     1081 gtgccctgtt catcggtgtg gctgtggccc cagctaacga catctacaac gagcgcgagc
     1141 tgctgaacag catgggcatc agccagccca ccgtcgtatt cgtgagcaag aaagggctgc
     1201 aaaagatcct caacgtgcaa aagaagctac cgatcataca aaagatcatc atcatggata
     1261 gcaagaccga ctaccagggc ttccaaagca tgtacacctt cgtgacttcc catttgccac
     1321 ccggcttcaa cgagtacgac ttcgtgcccg agagcttcga ccgggacaaa accatcgccc
     1381 tgatcatgaa cagtagtggc agtaccggat tgcccaaggg cgtagcccta ccgcaccgca
     1441 ccgcttgtgt ccgattcagt catgcccgcg accccatctt cggcaaccag atcatccccg
     1501 acaccgctat cctcagcgtg gtgccatttc accacggctt cggcatgttc accacgctgg
     1561 gctacttgat ctgcggcttt cgggtcgtgc tcatgtaccg cttcgaggag gagctattct
     1621 tgcgcagctt gcaagactat aagattcaat ctgccctgct ggtgcccaca ctatttagct
     1681 tcttcgctaa gagcactctc atcgacaagt acgacctaag caacttgcac gagatcgcca
     1741 gcggcggggc gccgctcagc aaggaggtag gtgaggccgt ggccaaacgc ttccacctac
     1801 caggcatccg ccagggctac ggcctgacag aaacaaccag cgccattctg atcacccccg
     1861 aaggggacga caagcctggc gcagtaggca aggtggtgcc cttcttcgag gctaaggtgg
     1921 tggacttgga caccggtaag acactgggtg tgaaccagcg cggcgagctg tgcgtccgtg
     1981 gccccatgat catgagcggc tacgttaaca accccgaggc tacaaacgct ctcatcgaca
     2041 aggacggctg gctgcacagc ggcgacatcg cctactggga cgaggacgag cacttcttca
     2101 tcgtggaccg gctgaagagc ctgatcaaat acaagggcta ccaggtagcc ccagccgaac
     2161 tggagagcat cctgctgcaa caccccaaca tcttcgacgc cggggtcgcc ggcctgcccg
     2221 acgacgatgc cggcgagctg cccgccgcag tcgtcgtgct ggaacacggt aaaaccatga
     2281 ccgagaagga gatcgtggac tatgtggcca gccaggttac aaccgccaag aagctgcgcg
     2341 gtggtgttgt gttcgtggac gaggtgccta aaggactgac cggcaagttg gacgcccgca
     2401 agatccgcga gattctcatt aaggccaaga agggcggcaa gatcgccgtg taagaattcg
     2461 cttatgcggc cgcctccttt aagaaggaga tatagatatc ccaactccat aagtgtgtcg
     2521 acatataagg atccactcga cgactcgaca atcaacctct ggattacaaa atttgtgaaa
     2581 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2641 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2701 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     2761 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     2821 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     2881 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     2941 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     3001 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3061 tgccggctct gcggcctctt ccgcatcttc gccttcgccc tcagacgagt cggatctccc
     3121 tttgggccgc ctccccgcct ggaattaatt cgagctcggt accttgattc tcgagtctag
     3181 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3241 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3301 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3361 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3421 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     3481 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3541 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3601 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3661 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3721 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3781 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3841 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3901 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3961 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4021 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4081 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4141 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4201 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4261 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     4321 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4381 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4441 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4501 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4561 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4621 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4681 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4741 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4801 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4861 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4921 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4981 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5041 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5101 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5161 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5221 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5281 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5341 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5401 atgtatctta tcatgtctgt ataccctcca cctctagcta gagcttggcg taatcatggt
     5461 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5521 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5581 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5641 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5701 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5761 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5821 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5881 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5941 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6001 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6061 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6121 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6181 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6241 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6301 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6361 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6421 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6481 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6541 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6601 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6661 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6721 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6781 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6841 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6901 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6961 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7021 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7081 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7141 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7201 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7261 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7321 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7381 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7441 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7501 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7561 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacctccacg
     7621 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
     7681 atagttaagc cagtatctgc tccctgcttg tgtgttggag gt
//
