LOCUS       sequence-403387-        7398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for mammalian cells: Expression of mScarlet 
            with a CMV 5'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7398)
  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 7398)
  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-Control-5'UTR
FEATURES             Location/Qualifiers
     source          1..7398
                     /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"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             597..1292
                     /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(730..748)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     998..1017
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1302..1890
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1355..1375)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1773..1784)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1977..2210
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2282..2403
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2319..2338)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2373..2392
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2443..2578
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2598..2617)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2599..2617)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2627..2644)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2627..2643)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2636..2658)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2785..3240
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2872..2891)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3082..3103
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3266..3370
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3371..3439,3440..4231)
                     /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(3589..3608)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4402..4990
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4891..4910
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5144..5161
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5278..5299
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5314..5319,5320..5337,5338..5344)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5352..5368
                     /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     5357..5379
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5376..5392
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5376..5392
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5411..5431
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5413..5431
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5459..5685
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5686..5866
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5913..6038
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6531..6764
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6949..6993
                     /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    7260..7376
                     /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 cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgactc tagaggatcc accggtatgg
      601 tgagcaaggg cgaggcagtg atcaaggagt tcatgcggtt caaggtgcac atggagggct
      661 ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
      721 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttctcc tgggacatcc
      781 tgtcccctca gttcatgtac ggctccaggg ccttcaccaa gcaccccgcc gacatccccg
      841 actactataa gcagtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
      901 acggcggcgc cgtgaccgtg acccaggaca cctccctgga ggacggcacc ctgatctaca
      961 aggtgaagct ccgcggcacc aacttccctc ctgacggccc cgtaatgcag aagaagacaa
     1021 tgggctggga agcgtccacc gagcggttgt accccgagga cggcgtgctg aagggcgaca
     1081 ttaagatggc cctgcgcctg aaggacggcg gccgctacct ggcggacttc aagaccacct
     1141 acaaggccaa gaagcccgtg cagatgcccg gcgcctacaa cgtcgaccgc aagttggaca
     1201 tcacctccca caacgaggac tacaccgtgg tggaacagta cgaacgctcc gagggccgcc
     1261 actccaccgg cggcatggac gagctgtaca agtgagtcga caatcaacct ctggattaca
     1321 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     1381 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     1441 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     1501 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     1561 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     1621 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     1681 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     1741 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     1801 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     1861 gtcggatctc cctttgggcc gcctccccgc ctggaattcg agctcggtac ctttaagacc
     1921 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     1981 agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg
     2041 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     2101 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     2161 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     2221 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     2281 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2341 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2401 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg
     2461 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2521 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2581 gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     2641 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     2701 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     2761 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2821 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2881 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2941 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3001 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3061 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3121 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3181 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     3241 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3301 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3361 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3421 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3481 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3541 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3601 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3661 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3721 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3781 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3841 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3901 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3961 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4021 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4081 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4141 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4201 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4261 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4321 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4381 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4441 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4501 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4561 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4621 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4681 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4741 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4801 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4861 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4921 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4981 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5041 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     5101 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     5161 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     5221 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     5281 tgtgagttag ctcactcatt aggcacccca ggctttccac tttatgcttc cggctcgtat
     5341 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     5401 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
     5461 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     5521 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     5581 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     5641 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     5701 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     5761 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     5821 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     5881 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     5941 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     6001 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     6061 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     6121 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     6181 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     6241 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     6301 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     6361 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     6421 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     6481 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     6541 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     6601 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     6661 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     6721 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     6781 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     6841 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     6901 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     6961 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     7021 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     7081 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     7141 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     7201 gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact
     7261 tttaaaagaa aaggggggat tggggggtat agtgcagggg aaagaatagt agacataata
     7321 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaattcaa aattttatcg
     7381 ataagcttgg gagttccg
//
