LOCUS       sequence-226800-        8814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express Raptor-Halo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8814)
  AUTHORS   Lawrence RE, Cho KF, Rappold R, Thrun A, Tofaute M, Kim DJ, 
            Moldavski O, Hurley JH, Zoncu R
  TITLE     A nutrient-induced affinity switch controls mTORC1 activation by its
            Rag GTPase-Ragulator lysosomal scaffold.
  JOURNAL   Nat Cell Biol. 2018 Sep;20(9):1052-1063. doi: 
            10.1038/s41556-018-0148-6. Epub 2018 Jul 30.
  PUBMED    30061680
REFERENCE   2  (bases 1 to 8814)
  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: 
            "10.1038/s41556-018-0148-6"; journalName: "Nat Cell Biol"; date: 
            "2018-09"; volume: "20"; issue: "9"; pages: "1052-1063"
COMMENT     Sequence Label: pEGFP-N1 Raptor Halo
FEATURES             Location/Qualifiers
     source          1..8814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      619..628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4663..5550
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     polyA_signal    5675..5796
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5712..5731)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5766..5785
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5803..6257)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5939..5960)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6151..6170
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6284..6388
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6413..6433)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6457..6667
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        6479..6675
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6526..6661
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6710..7504
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6710..7504
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6764..6783)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7374..7393
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7692..7711)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7736..7783
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8112..8700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8601..8620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgaggc caccatggag tccgaaatgc tgcaatcgcc tcttctgggc
      661 ctgggggagg aagatgaggc tgatcttaca gactggaacc tacctttggc ttttatgaaa
      721 aagaggcact gtgagaaaat tgaaggctcc aaatccttag ctcagagctg gaggatgaag
      781 gatcggatga agacagtcag tgttgcctta gttttgtgcc tgaatgttgg tgtggaccct
      841 cccgatgtgg tgaagaccac gccctgtgca cgcttggaat gctggatcga tcctctgtcg
      901 atgggtcctc agaaagctct ggaaaccatc ggtgcaaatt tacagaagca gtacgagaac
      961 tggcagccaa gggcccggta caagcagagc cttgacccaa ctgtggatga agtcaagaag
     1021 ctctgcacgt ccttacgtcg caacgccaag gaggagcgag tcctctttca ctacaatggc
     1081 cacggggtgc cccggcccac agtcaacggg gaggtctggg tcttcaacaa gaactacacg
     1141 cagtacatcc ctctgtccat atatgacctg cagacgtgga tgggcagccc gtcgatcttc
     1201 gtctacgact gctccaatgc tggcttgatc gtcaagtcct tcaagcagtt cgcactacag
     1261 cgggagcagg agctggaggt agctgcaatc aacccaaatc accctcttgc tcagatgcct
     1321 ttgcctccgt cgatgaaaaa ctgcatccag ctggcagcct gcgaggccac cgagctgctg
     1381 cccatgatcc ccgacctccc ggctgaccta ttcacctcct gcctcaccac ccccatcaag
     1441 atcgccctgc gctggttttg catgcagaaa tgtgtcagtc tggtgcctgg cgtcacactg
     1501 gatttgatag aaaagatccc tggccgcctg aacgacagga ggacgcccct gggtgaactg
     1561 aactggatct tcacagccat cacagacacc atcgcgtgga acgtgctccc ccgggatctc
     1621 ttccaaaagc tcttcagaca ggacttgctg gtggctagtc tgtttcgaaa ttttttattg
     1681 gcggaaagga ttatgaggtc gtataactgc actcccgtca gcagcccgcg tctgccgccc
     1741 acgtacatgc acgccatgtg gcaagcctgg gacctggctg ttgacatctg tctgtctcag
     1801 ctgccgacga tcatcgagga aggcactgcg tttcggcaca gcccgttctt cgccgagcag
     1861 ctgaccgcat tccaggtgtg gctcaccatg ggcgtggaga accgaaaccc acccgaacag
     1921 ctgcccatcg tcctgcaggt gctgttaagc caagtgcacc ggctgagagc attggacttg
     1981 cttggaagat ttttggacct gggtccctgg gcagtgagcc tggccttgtc tgtcggcatc
     2041 ttcccctacg tgctgaagct gctccagagc tcggcccgag agctgcggcc acttctcgtt
     2101 ttcatctggg ccaagatcct cgcagtggac agctcgtgcc aggcggacct cgtgaaggac
     2161 aacggccaca agtacttcct gtcggtcctg gcggacccct acatgccagc tgaacaccgg
     2221 accatgacgg ctttcattct cgccgtgatc gtcaacagct atcacacggg gcaggaagcc
     2281 tgccttcagg gaaacctcat tgccatctgc ctggagcagc tcaacgaccc gcaccccttg
     2341 ctgcgccagt gggtggccat ctgcctcggc aggatctggc agaacttcga ctcggcgagg
     2401 tggtgcggcg tgagggacag cgctcatgag aagctctaca gcctcctctc cgaccccatt
     2461 cccgaggtcc gctgcgcagc ggtcttcgcc cttggcacgt tcgtgggcaa ctctgcagag
     2521 aggacggacc actccaccac catcgaccac aacgtggcca tgatgctggc ccagctggtc
     2581 agcgacggga gccccatggt ccggaaggag ctggtggtgg ctctgagtca tcttgtggtt
     2641 cagtatgaaa gcaatttctg caccgtggcc ctgcagttca tagaagagga aaagaactac
     2701 gccttgcctt ctccagcaac cacagaggga gggagtttga ccccagtgcg agacagcccg
     2761 tgcaccccca gacttcgttc tgtgagctcc tatggaaaca tccgtgctgt cgccacagcc
     2821 aggagcctca acaaatcttt gcagaacctg agtttgacag aggaatctgg tggcgcggtg
     2881 gcgttctccc ccggaaacct cagcaccagc agcagcgcca gcagcaccct gggcagcccc
     2941 gagaatgagg agcatatcct gtccttcgag accatcgaca agatgcgccg cgccagctcc
     3001 tactcctccc tcaactccct catcggagtt tcctttaaca gtgtttacac tcagatttgg
     3061 agagtcctgc tgcacctggc tgctgacccc tatccagagg tctcggacgt ggccatgaaa
     3121 gtactcaaca gcatcgccta caaggccacc gtgaacgccc ggccgcagcg cgtcctggac
     3181 acctcctccc tcacgcagtc ggcccccgcc agccccacca acaagggcgt gcacatccac
     3241 caggcggggg gctcccctcc ggcgtccagc accagcagct ccagcctgac caacgatgtg
     3301 gccaagcagc cggtcagccg agacttgcct tctggccggc cgggcaccac aggccccgct
     3361 ggggcgcagt acacccctca ctcccaccag ttcccccgga cacggaagat gttcgacaag
     3421 ggcccagagc agactgcgga cgacgcggac gatgctgctg gacacaaaag tttcatctcc
     3481 gccacggtgc agacggggtt ctgcgactgg agcgcccgct attttgccca gcccgtcatg
     3541 aagatcccag aagagcacga cctggagagt cagatccgca aggagcggga gtggcggttc
     3601 ctgcgaaaca gccgtgtcag gaggcaggcc cagcaagtca ttcagaaggg cattacgaga
     3661 ttggacgacc aaatatttct gaacaggaac cccggcgtcc cctctgtggt gaaattccac
     3721 cccttcacgc cgtgcatcgc tgtagccgac aaggacagca tctgcttttg ggactgggag
     3781 aaaggggaga agctggatta tttccacaat gggaaccctc ggtacacgag ggtcactgcc
     3841 atggagtatc tgaatggcca ggactgctcg cttctgctga cggccacaga cgatggtgcc
     3901 atcagggtct ggaagaattt tgctgatttg gaaaagaacc cagagatggt gaccgcgtgg
     3961 caggggctct cggacatgct gccaacgacg cgaggagctg ggatggtggt ggactgggag
     4021 caggagaccg gcctcctcat gagctcagga gacgtgcgga tcgtccggat ctgggacaca
     4081 gaccgtgaga tgaaggtgca ggacatccct acgggcgcag acagctgtgt gacgagtctg
     4141 tcctgtgatt cccaccgctc actcatcgtg gctggcctcg gtgacggctc catccgcgtc
     4201 tacgacagaa ggatggcact cagcgaatgc cgcgtcatga cgtaccggga gcacacagcc
     4261 tgggtggtga aggcctccct gcagaagcgt cccgacggcc acatcgtgag tgtgagcgtc
     4321 aatggagatg tgcgcatctt tgatccccgg atgcctgagt cggtaaatgt gcttcagatc
     4381 gtgaaggggc tgacggccct ggacatccac ccccaggcgg acctgatcgc atgtggctcc
     4441 gtcaatcagt tcaccgccat ctacaacagc agcggagagc tcatcaacaa catcaagtac
     4501 tacgacggct tcatgggcca gcgggtcggc gccatcagct gcctggcctt ccacccgcac
     4561 tggcctcacc tggccgtggg aagcaacgac tactacatct ccgtgtactc ggtggagaag
     4621 cgtgtcagac gaattctgca gtcgacgggt ggtggtggtg gtatggaaat cggtactggc
     4681 tttccattcg acccccatta tgtggaagtc ctgggcgagc gcatgcacta cgtagatgtt
     4741 ggtccgcgcg atggcacccc tgtgctgttc ctgcacggta acccgacctc ctcctacgtg
     4801 tggcgcaaca tcatcccgca tgttgcaccg acccatcgct gcattgctcc agacctgatc
     4861 ggtatgggca aatccgacaa accagacctg ggttatttct tcgacgacca cgtccgcttc
     4921 atggatgcct tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat tcacgactgg
     4981 ggctccgctc tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa aggtattgca
     5041 tttatggagt tcatccgccc tatcccgacc tgggacgaat ggccagaatt tgcccgcgag
     5101 accttccagg ccttccgcac caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt
     5161 tttatcgagg gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt cgagatggac
     5221 cattaccgcg agccgttcct gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac
     5281 gagctgccaa tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga atacatggac
     5341 tggctgcacc agtcccctgt cccgaagctg ctgttctggg gcaccccagg cgttctgatc
     5401 ccaccggccg aagccgctcg cctggccaaa agcctgccta actgcaaggc tgtggacatc
     5461 ggcccgggtc tgaatctgct gcaagaagac aacccggacc tgatcggcag cgagatcgcg
     5521 cgctggctgt ctactctgga gatttccggt taggcggccg cgactctaga tcataatcag
     5581 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     5641 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     5701 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5761 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     5821 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac aataggccga
     5881 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     5941 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     6001 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     6061 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     6121 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     6181 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     6241 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6301 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6361 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     6421 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     6481 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     6541 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     6601 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     6661 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     6721 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     6781 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     6841 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     6901 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     6961 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     7021 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     7081 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     7141 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     7201 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     7261 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     7321 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     7381 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     7441 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     7501 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     7561 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     7621 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     7681 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     7741 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     7801 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     7861 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     7921 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     7981 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     8041 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     8101 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     8161 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     8221 ataactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     8281 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     8341 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     8401 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     8461 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     8521 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     8581 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     8641 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     8701 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     8761 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
