LOCUS       sequence-392071-        7014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of LYCHOS (GPR155) ∆DEP-HA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7014)
  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 7014)
  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) ∆DEP-HA
FEATURES             Location/Qualifiers
     source          1..7014
                     /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             3270..3296
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3314..3323
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3327..3461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3364..3383)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3418..3437
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3530..3887
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3530..3721
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3553..3573)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3738..3873
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3907..3924)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3907..3923)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3916..3938)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4136..4591
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4223..4242)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4433..4454
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4608..4627)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4727..4749
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4787..4805)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4873..4977
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4978..5046,5047..5838)
                     /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(5196..5215)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6009..6597
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6498..6517
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6751..6768
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6885..6906
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6921..6926,6927..6944,6945..6951)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6959..6975
                     /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     6964..6986
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6983..6999
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6983..6999
                     /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 ttcctcctgc tattaatgca aacactctcc
     3181 aacaggaaag atataaagaa attgagcatt catccccacc ctcacattcc cctaagacca
     3241 gatctaatgg tggtggtggt gggatgggct acccctatga tgtgcctgac tacgcataag
     3301 gatccccagc ttggccgcca tggcccaact tgtttattgc agcttataat ggttacaaat
     3361 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3421 gtttgtccaa actcatcaat gtatcttatc atgtctggat cgggaattaa ttcggcgcag
     3481 caccatggcc tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa
     3541 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3601 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3661 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3721 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3781 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3841 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctgttaacag
     3901 cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     3961 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     4021 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt
     4081 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     4141 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4201 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4261 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4321 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     4381 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4441 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg
     4501 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4561 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     4621 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4681 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4741 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4801 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4861 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4921 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4981 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5041 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5101 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5161 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5221 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5281 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5341 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5401 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5461 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5521 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5581 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5641 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5701 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5761 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5821 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5881 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5941 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6001 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6061 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6121 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6181 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6241 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6301 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6361 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6421 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6481 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6541 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6601 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6661 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6721 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6781 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     6841 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     6901 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     6961 gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa ttaa
//
