LOCUS       sequence-363197-        7457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for mammalian cells: Expression of mScarlet 
            with MAVS 5'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7457)
  AUTHORS   Vora SM, Fontana P, Mao T, Leger V, Zhang Y, Fu TM, Lieberman J, 
            Gehrke L, Shi M, Wang L, Iwasaki A, Wu H
  TITLE     Targeting stem-loop 1 of the SARS-CoV-2 5' UTR to suppress viral 
            translation and Nsp1 evasion.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 Mar 1;119(9). pii: 2117198119. doi: 
            10.1073/pnas.2117198119.
  PUBMED    35149555
REFERENCE   2  (bases 1 to 7457)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2022-03-1"; volume: "119(9). pii: 
            2117198119. doi"; pages: " 10.1073/pnas.2117198119"
COMMENT     Sequence Label: pLV-mScarlet-MAVS-5'UTR
FEATURES             Location/Qualifiers
     source          1..7457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             656..1351
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(789..807)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1057..1076
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1361..1949
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1414..1434)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1832..1843)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2036..2269
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2341..2462
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2378..2397)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2432..2451
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2502..2637
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2657..2676)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2658..2676)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2686..2703)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2686..2702)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2695..2717)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2844..3299
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2931..2950)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3141..3162
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3325..3429
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3430..3498,3499..4290)
                     /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(3648..3667)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4461..5049
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4950..4969
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5203..5220
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5337..5358
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5373..5378,5379..5396,5397..5403)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5411..5427
                     /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)."
     primer_bind     5416..5438
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5435..5451
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5435..5451
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5470..5490
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5472..5490
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5518..5744
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5745..5925
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5972..6097
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6590..6823
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7008..7052
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7319..7435
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac caagcttgcg ggaagggtcc
      541 tgggccccgg gcggcggtcg ccaggtctca gggccggggg tacccgagtc tcgtttcctc
      601 tcagtccatc cacccttcat ggggccagag ccctctctcc agaatctgag cagcaatggt
      661 gagcaagggc gaggcagtga tcaaggagtt catgcggttc aaggtgcaca tggagggctc
      721 catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
      781 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttctcct gggacatcct
      841 gtcccctcag ttcatgtacg gctccagggc cttcaccaag caccccgccg acatccccga
      901 ctactataag cagtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
      961 cggcggcgcc gtgaccgtga cccaggacac ctccctggag gacggcaccc tgatctacaa
     1021 ggtgaagctc cgcggcacca acttccctcc tgacggcccc gtaatgcaga agaagacaat
     1081 gggctgggaa gcgtccaccg agcggttgta ccccgaggac ggcgtgctga agggcgacat
     1141 taagatggcc ctgcgcctga aggacggcgg ccgctacctg gcggacttca agaccaccta
     1201 caaggccaag aagcccgtgc agatgcccgg cgcctacaac gtcgaccgca agttggacat
     1261 cacctcccac aacgaggact acaccgtggt ggaacagtac gaacgctccg agggccgcca
     1321 ctccaccggc ggcatggacg agctgtacaa gtgagtcgac aatcaacctc tggattacaa
     1381 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     1441 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     1501 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     1561 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     1621 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     1681 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     1741 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     1801 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     1861 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     1921 tcggatctcc ctttgggccg cctccccgcc tggaattcga gctcggtacc tttaagacca
     1981 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     2041 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
     2101 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     2161 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     2221 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     2281 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     2341 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2401 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2461 tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc
     2521 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2581 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2641 ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     2701 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     2761 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2821 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2881 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2941 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3001 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3061 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3121 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3181 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3241 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     3301 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3361 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3421 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3481 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3541 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3601 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3661 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3721 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3781 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3841 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3901 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3961 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4021 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4081 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4141 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4201 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4261 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4321 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4381 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4441 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4501 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4561 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4621 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4681 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4741 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4801 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4861 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4921 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4981 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5041 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5101 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5161 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5221 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     5281 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     5341 gtgagttagc tcactcatta ggcaccccag gctttccact ttatgcttcc ggctcgtatg
     5401 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     5461 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt
     5521 agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct
     5581 tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg
     5641 ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc
     5701 attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag
     5761 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     5821 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     5881 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
     5941 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
     6001 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     6061 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     6121 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     6181 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     6241 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     6301 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     6361 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     6421 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     6481 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     6541 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     6601 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
     6661 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     6721 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     6781 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     6841 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     6901 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
     6961 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
     7021 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
     7081 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
     7141 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
     7201 cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag
     7261 agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat cggttaactt
     7321 ttaaaagaaa aggggggatt ggggggtata gtgcagggga aagaatagta gacataatag
     7381 caacagacat acaaactaaa gaattacaaa aacaaattac aaaattcaaa attttatcga
     7441 taagcttggg agttccg
//
