LOCUS       sequence-392073-        7321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of LYCHOS (GPR155) WT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7321)
  AUTHORS   Shin HR, Citron YR, Wang L, Tribouillard L, Goul CS, Stipp R, 
            Sugasawa Y, Jain A, Samson N, Lim CY, Davis OB, Castaneda-Carpio D, 
            Qian M, Nomura DK, Perera RM, Park E, Covey DF, Laplante M, Evers 
            AS, Zoncu R
  TITLE     Lysosomal GPCR-like protein LYCHOS signals cholesterol sufficiency 
            to mTORC1.
  JOURNAL   Science. 2022 Sep 16;377(6612):1290-1298. doi: 
            10.1126/science.abg6621. Epub 2022 Aug 25.
  PUBMED    36007018
REFERENCE   2  (bases 1 to 7321)
  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.abg6621"; journalName: "Science"; date: 
            "2022-09-16- 16"; volume: "377"; issue: "6612"; pages: "1290-1298"
COMMENT     Sequence Label: pRK5-LYCHOS (GPR155) WT-myc
FEATURES             Location/Qualifiers
     source          1..7321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             complement(2819..2830)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             3555..3584
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3621..3630
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3634..3768
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3671..3690)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3725..3744
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3837..4194
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3837..4028
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3860..3880)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4045..4180
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4214..4231)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4214..4230)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4223..4245)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4443..4898
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4530..4549)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4740..4761
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4915..4934)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5034..5056
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5094..5112)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5180..5284
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5285..5353,5354..6145)
                     /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(5503..5522)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6316..6904
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6805..6824
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7058..7075
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7192..7213
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7228..7233,7234..7251,7252..7258)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7266..7282
                     /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     7271..7293
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7290..7306
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7290..7306
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgatgcc accatgaatt ctaatttacc tgcagagaac ttaaccattg
      961 cagtcaatat gaccaagact ttgcctacag cagtaacgca tggatttaat tccactaatg
     1021 acccaccttc aatgtcaatt acaaggcttt ttccagcctt actggaatgc tttggcattg
     1081 tcctttgtgg ctacatagca ggaagggcca atgtcataac atcaacccag gccaaaggac
     1141 taggaaattt tgtctccaga tttgcacttc cagctttatt attcaaaaac atggttgtac
     1201 ttaatttttc caatgtggac tggtccttcc tatatagtat cttaattgcc aaagcttctg
     1261 tatttttcat tgtatgtgta ttaaccttat tggttgccag tcctgatagt cgatttagca
     1321 aagctggact attccctatt tttgctacac aaagtaatga ctttgcattg ggatacccta
     1381 tagttgaagc tttatatcaa actacatacc cagaatatct ccagtacatt tatttggtgg
     1441 caccaatatc tcttatgatg ttaaacccta tagggtttat tttctgtgaa atccaaaagt
     1501 ggaaagacac tcaaaatgct tctcaaaata aaataaaaat tgtgggactc ggactcctgc
     1561 gtgtattaca gaacccaata gtatttatgg tcttcattgg catcgccttc aattttattc
     1621 ttgatcgaaa ggtacctgta tatgtcgaaa attttcttga tggacttgga aattcttttt
     1681 ctggatcagc cctattttat cttggtctca cgatggtggg aaaaataaag agactgaaga
     1741 agtcggcatt tgtagtacta attcttctca tcacagctaa acttctggtg ctgccacttc
     1801 tgtgcagaga aatggtggaa ctcttggaca agggcgacag tgtggtgaac catacaagtt
     1861 tatcaaatta tgcatttctg tatggtgtat ttcctgtagc accaggagtg gctatctttg
     1921 caacacaatt caacatggaa gtagaaatta taacctcagg gatggtgata agcacatttg
     1981 tgtctgctcc catcatgtac gtttctgcct ggttactgac ctttcccact atggacccta
     2041 agccattggc atatgccatc cagaatgtta gttttgatat aagtattgtc agcctgatct
     2101 ccttgatctg gtctctggct attcttcttt tgagtaagaa atataaacag cttcctcata
     2161 tgcttacaac taatttactc attgctcagt ctattgtctg tgctggaatg atgatatgga
     2221 attttgttaa agaaaaaaat tttgttggac aaattttggt gtttgttcta ttgtacagct
     2281 ccctctatag cacctacctg tggacaggcc ttctagcaat ttctttgttt cttttgaaaa
     2341 agcgagagag ggtacaaatt cctgttggaa taatcataat atctggctgg ggaattcctg
     2401 ctctccttgt tggtgttctt ttgataactg gaaaacacaa tggagatagc attgactcag
     2461 ccttctttta tggaaaagaa cagatgatca ccacagcagt caccctgttc tgcagcatcc
     2521 tgatagctgg catatccctc atgtgcatga accagactgc ccaagcagga agctatgaag
     2581 gtttcgatca gtctcagagc cacaaagtgg tggagcctgg aaatactgct tttgaggaga
     2641 gtccagcacc agtaaatgaa ccagaacttt ttacaagctc tattccagaa acaagttgct
     2701 gctcctgctc catgggaaat ggtgaattac actgcccatc aatagagcca atagcaaaca
     2761 caagcaccag tgagcctgtg attccttcgt ttgagaaaaa caatcattgt gtgagtcgct
     2821 gtaactccca gagctgcata ttagcccagg aagaagaaca gtatctacag agtggagacc
     2881 agcaactgac ccgacatgtg ttgctgtgtt tacttctcat cattggcctg ttcgctaatc
     2941 tttccagttg tttatggtgg ctattcaacc aagagcctgg aagactttat gttgagttac
     3001 agtttttctg tgccgtgttt aactttggtc agggatttat ttcctttgga atctttggat
     3061 tagataaaca tttaatcatc ctgcctttca aaagaagact tgaattccta tggaacaata
     3121 aagacacagc agaaaacagg gattctcctg tttcagagga aataaaaatg acctgtcaac
     3181 aatttatcca ttatcaccgt gacctctgta tccgaaacat tgtcaaagaa agaaggtgtg
     3241 gtgcaaagac ttctgctgga actttctgtg gctgtgacct ggtgagctgg ctaattgaag
     3301 tcggccttgc ctccgaccgt ggtgaagctg tgatatacgg agacaggctg gtacaagggg
     3361 gagtcatcca acatattacc aatgagtatg aattccggga tgagtacttg ttttacagat
     3421 ttcttcaaaa gagtcctgaa cagagtcctc ctgctattaa tgcaaacact ctccaacagg
     3481 aaagatataa agaaattgag cattcatccc caccctcaca ttcccctaag accgtcgacg
     3541 gtggtggtgg tggtgaacaa aaactcatct cagaagagga tctgtaagcg gccgctaagt
     3601 aagtaaggat ccccagcttg gccgccatgg cccaacttgt ttattgcagc ttataatggt
     3661 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3721 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcgg gaattaattc
     3781 ggcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg taccttctga
     3841 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3901 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3961 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4021 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4081 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4141 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctg
     4201 ttaacagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     4261 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     4321 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     4381 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     4441 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4501 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4561 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4621 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     4681 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4741 tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt
     4801 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4861 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     4921 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4981 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     5041 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     5101 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     5161 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5221 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5281 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5341 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5401 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5461 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5521 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5581 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5641 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5701 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5761 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5821 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5881 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5941 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6001 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6061 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6121 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6181 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6241 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6301 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6361 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6421 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6481 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6541 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6601 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6661 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6721 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6781 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6841 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6901 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6961 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     7021 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     7081 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     7141 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     7201 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     7261 tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga ttacgaatta
     7321 a
//
