LOCUS       sequence-393289-        8239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of LYCHOS FP>IA mutant in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8239)
  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 8239)
  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: pFB-CG-LYCHOS FP>IA-HA-GFP
FEATURES             Location/Qualifiers
     source          1..8239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..557
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(189..208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     399..420
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        583..687
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(688..756,757..1548)
                     /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(906..925)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1719..2307
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2208..2227
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2461..2478
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2612..2836
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2903..3436)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3625..3630,3631..3647,3648..3653))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        4005..4096
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4021..4038
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             complement(6040..6051)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             6770..6787
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6857..6883
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6893..6916
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             7664..7693
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     polyA_signal    7829..7963
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7866..7885)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7920..7939
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7992..8157
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
      421 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
      481 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
      541 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
      601 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
      661 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
      721 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
      781 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
      841 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
      901 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
      961 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     1021 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     1081 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     1141 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     1201 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     1261 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     1321 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     1381 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     1441 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     1501 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     1561 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     1621 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1681 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1741 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1801 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1861 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1921 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1981 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2041 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2101 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2161 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2221 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2281 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2341 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2401 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     2461 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     2521 ctgtgcggta tttcacaccg catagaccag ccgcgtaacc tggcaaaatc ggttacggtt
     2581 gagtaataaa tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga
     2641 acaaaataga tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg
     2701 aaatcagtcc agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac
     2761 tcttcatttt ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ccaagggcat
     2821 ggtaaagact atattcgcgg cgttgtgaca atttaccgaa caactccgcg gccgggaagc
     2881 cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca aagtgcatca
     2941 cttcttcccg tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg
     3001 cttgcacgta gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga
     3061 gcgcggtggc aatgccctgc ctccggtgct cgccggagac tgcgagatca tagatataga
     3121 tctcactacg cggctgctca aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg
     3181 cttcttggtc gaaggcagca agcgcgatga atgtcttact acggagcaag ttcccgaggt
     3241 aatcggagtc cggctgatgt tgggagtagg tggctacgtc tccgaactca cgaccgaaaa
     3301 gatcaagagc agcccgcatg gatttgactt ggtcagggcc gagcctacat gtgcgaatga
     3361 tgcccatact tgagccacct aactttgttt tagggcgact gccctgctgc gtaacatcgt
     3421 tgctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
     3481 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg
     3541 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     3601 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt
     3661 cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc
     3721 gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag
     3781 gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct gccctggctt
     3841 caggagatcg gaagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa
     3901 gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc
     3961 tatagttcta gtggttggct acgtatactc cggaatatta atagatcatg gagataatta
     4021 aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat
     4081 aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc gcggatccgc
     4141 caccatgaat tctaatttac ctgcagagaa cttaaccatt gcagtcaata tgaccaagac
     4201 tttgcctaca gcagtaacgc atggatttaa ttccactaat gacccacctt caatgtcaat
     4261 tacaaggctt attgcagcct tactggaatg ctttggcatt gtcctttgtg gctacatagc
     4321 aggaagggcc aatgtcataa catcaaccca ggccaaagga ctaggaaatt ttgtctccag
     4381 atttgcactt ccagctttat tattcaaaaa catggttgta cttaattttt ccaatgtgga
     4441 ctggtccttc ctatatagta tcttaattgc caaagcttct gtatttttca ttgtatgtgt
     4501 attaacctta ttggttgcca gtcctgatag tcgatttagc aaagctggac tattccctat
     4561 ttttgctaca caaagtaatg actttgcatt gggataccct atagttgaag ctttatatca
     4621 aactacatac ccagaatatc tccagtacat ttatttggtg gcaccaatat ctcttatgat
     4681 gttaaaccct atagggttta ttttctgtga aatccaaaag tggaaagaca ctcaaaatgc
     4741 ttctcaaaat aaaataaaaa ttgtgggact cggactcctg cgtgtattac agaacccaat
     4801 agtatttatg gtcttcattg gcatcgcctt caattttatt cttgatcgaa aggtacctgt
     4861 atatgtcgaa aattttcttg atggacttgg aaattctttt tctggatcag ccctatttta
     4921 tcttggtctc acgatggtgg gaaaaataaa gagactgaag aagtcggcat ttgtagtact
     4981 aattcttctc atcacagcta aacttctggt gctgccactt ctgtgcagag aaatggtgga
     5041 actcttggac aagggcgaca gtgtggtgaa ccatacaagt ttatcaaatt atgcatttct
     5101 gtatggtgta tttcctgtag caccaggagt ggctatcttt gcaacacaat tcaacatgga
     5161 agtagaaatt ataacctcag ggatggtgat aagcacattt gtgtctgctc ccatcatgta
     5221 cgtttctgcc tggttactga cctttcccac tatggaccct aagccattgg catatgccat
     5281 ccagaatgtt agttttgata taagtattgt cagcctgatc tccttgatct ggtctctggc
     5341 tattcttctt ttgagtaaga aatataaaca gcttcctcat atgcttacaa ctaatttact
     5401 cattgctcag tctattgtct gtgctggaat gatgatatgg aattttgtta aagaaaaaaa
     5461 ttttgttgga caaattttgg tgtttgttct attgtacagc tccctctata gcacctacct
     5521 gtggacaggc cttctagcaa tttctttgtt tcttttgaaa aagcgagaga gggtacaaat
     5581 tcctgttgga ataatcataa tatctggctg gggaattcct gctctccttg ttggtgttct
     5641 tttgataact ggaaaacaca atggagatag cattgactca gccttctttt atggaaaaga
     5701 acagatgatc accacagcag tcaccctgtt ctgcagcatc ctgatagctg gcatatccct
     5761 catgtgcatg aaccagactg cccaagcagg aagctatgaa ggtttcgatc agtctcagag
     5821 ccacaaagtg gtggagcctg gaaatactgc ttttgaggag agtccagcac cagtaaatga
     5881 accagaactt tttacaagct ctattccaga aacaagttgc tgctcctgct ccatgggaaa
     5941 tggtgaatta cactgcccat caatagagcc aatagcaaac acaagcacca gtgagcctgt
     6001 gattccttcg tttgagaaaa acaatcattg tgtgagtcgc tgtaactccc agagctgcat
     6061 attagcccag gaagaagaac agtatctaca gagtggagac cagcaactga cccgacatgt
     6121 gttgctgtgt ttacttctca tcattggcct gttcgctaat ctttccagtt gtttatggtg
     6181 gctattcaac caagagcctg gaagacttta tgttgagtta cagtttttct gtgccgtgtt
     6241 taactttggt cagggattta tttcctttgg aatctttgga ttagataaac atttaatcat
     6301 cctgcctttc aaaagaagac ttgaattcct atggaacaat aaagacacag cagaaaacag
     6361 ggattctcct gtttcagagg aaataaaaat gacctgtcaa caatttatcc attatcaccg
     6421 tgacctctgt atccgaaaca ttgtcaaaga aagaaggtgt ggtgcaaaga cttctgctgg
     6481 aactttctgt ggctgtgacc tggtgagctg gctaattgaa gtcggccttg cctccgaccg
     6541 tggtgaagct gtgatatacg gagacaggct ggtacaaggg ggagtcatcc aacatattac
     6601 caatgagtat gaattccggg atgagtactt gttttacaga tttcttcaaa agagtcctga
     6661 acagagtcct cctgctatta atgcaaacac tctccaacag gaaagatata aagaaattga
     6721 gcattcatcc ccaccctcac attcccctaa gaccggtggc tccggtggac atcaccacca
     6781 tcatcacgga tccggtggtt ctgccggtgg ctccggttct ggctccagcg gtggcagctc
     6841 tggtgcggcg gccgcatacc catacgatgt tccagattac gcttcgaatt ctctggaggt
     6901 gctgttccag ggtcctgcgg ccgccgtctc aaagggagaa gagctgttca ctggtgttgt
     6961 tccaatcctc gtggaactcg acggtgatgt caacggccat aagttcagcg tatccggaga
     7021 aggtgaaggc gacgcaacct acggtaaact gacactgaag tttatttgca ccaccggaaa
     7081 gctgccagtg ccatggccca ctttggtgac gacgttgact tacggagtcc aatgtttcag
     7141 ccgctacccg gatcacatga agcaacacga cttctttaag tctgctatgc ccgagggata
     7201 cgtccaggag cgcactattt tctttaaaga tgatggaaac tacaagacaa gagccgaggt
     7261 gaagttcgag ggtgacaccc tcgtgaaccg tattgagctt aagggcattg actttaagga
     7321 agatggcaac atcctgggac acaagctcga atacaactac aattcccata acgtatacat
     7381 catggcggac aaacaaaaga atggcatcaa ggtaaacttc aagatcagac ataacatcga
     7441 ggatggttcg gtgcaacttg cagaccacta ccagcagaac accccaatcg gtgacggtcc
     7501 cgtgctcctt cccgacaacc actacctgag cacacagtcc aagctatcaa aagaccctaa
     7561 cgaaaagcgt gaccacatgg tgcttctcga atttgtcaca gctgctggta tcaccctggg
     7621 tatggacgaa ctgtacaagt cgggactgcg tagtgcggcc gcacaccatc atcatcacca
     7681 ccatcaccat cactgataaa agcttgtcga gaagtactag aggatcataa tcagccatac
     7741 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     7801 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     7861 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     7921 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgatcac tgcttgagcc
     7981 taggagatcc gaaccagata agtgaaatct agttccaaac tattttgtca tttttaattt
     8041 tcgtattagc ttacgacgct acacccagtt cccatctatt ttgtcactct tccctaaata
     8101 atccttaaaa actccatttc cacccctccc agttcccaac tattttgtcc gcccacagcg
     8161 gggcattttt cttcctgtta tgtttttaat caaacatcct gccaactcca tgtgacaaac
     8221 cgtcatcttc ggctacttt
//
