LOCUS       sequence-125871-       10340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of human MTOR [NM_004958.3] without stop codon 
            (for C-terminal fusions).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10340)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 10340)
  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"
COMMENT     Sequence Label: R777-E138 Hs.MTOR-nostop
FEATURES             Location/Qualifiers
     source          1..10340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    84..183
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      3037..3046
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6246..6524
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     CDS             6258..6530
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQL"
     protein_bind    complement(7831..7930)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             8307..9098
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      9191..9779
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9680..9699
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9933..9950
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(10109..10136)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(10228..10314)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa
       61 attcgagctc ggtccgagga tccgaaataa tgattttatt ttgactgata gtgacctgtt
      121 cgttgcaaca aattgataag caatgctttt ttataatgcc aactttgtac aaaaaagttg
      181 gcaccatgct tggaaccgga cctgccgccg ccaccaccgc tgccaccaca tctagcaatg
      241 tgagcgtcct gcagcagttt gccagtggcc taaagagccg gaatgaggaa accagggcca
      301 aagccgccaa ggagctccag cactatgtca ccatggaact ccgagagatg agtcaagagg
      361 agtctactcg cttctatgac caactgaacc atcacatttt tgaattggtt tccagctcag
      421 atgccaatga gaggaaaggt ggcatcttgg ccatagctag cctcatagga gtggaaggtg
      481 ggaatgccac ccgaattggc agatttgcca actatcttcg gaacctcctc ccctccaatg
      541 acccagttgt catggaaatg gcatccaagg ccattggccg tcttgccatg gcaggggaca
      601 cttttaccgc tgagtacgtg gaatttgagg tgaagcgagc cctggaatgg ctgggtgctg
      661 accgcaatga gggccggaga catgcagctg tcctggttct ccgtgagctg gccatcagcg
      721 tccctacctt cttcttccag caagtgcaac ccttctttga caacattttt gtggccgtgt
      781 gggaccccaa acaggccatc cgtgagggag ctgtagccgc ccttcgtgcc tgtctgattc
      841 tcacaaccca gcgtgagccg aaggagatgc agaagcctca gtggtacagg cacacatttg
      901 aagaagcaga gaagggattt gatgagacct tggccaaaga gaagggcatg aatcgggatg
      961 atcggatcca tggagccttg ttgatcctta acgagctggt ccgaatcagc agcatggagg
     1021 gagagcgtct gagagaagaa atggaagaaa tcacacagca gcagctggta cacgacaagt
     1081 actgcaaaga tctcatgggc ttcggaacaa aacctcgtca cattaccccc ttcaccagtt
     1141 tccaggctgt acagccccag cagtcaaatg ccttggtggg gctgctgggg tacagctctc
     1201 accaaggcct catgggattt gggacctccc ccagtccagc taagtccacc ctggtggaga
     1261 gccggtgttg cagagacttg atggaggaga aatttgatca ggtgtgccag tgggtgctga
     1321 aatgcaggaa tagcaagaac tcgctgatcc aaatgacaat ccttaatttg ttgccccgct
     1381 tggctgcatt ccgaccttct gccttcacag atacccagta tctccaagat accatgaacc
     1441 atgtcctaag ctgtgtcaag aaggagaagg aacgtacagc ggccttccaa gccctggggc
     1501 tactttctgt ggctgtgagg tctgagttta aggtctattt gcctcgcgtg ctggacatca
     1561 tccgagcggc cctgccccca aaggacttcg cccataagag gcagaaggca atgcaggtgg
     1621 acgccacagt cttcacttgc atcagcatgc tggctcgagc aatggggcca ggcatccagc
     1681 aggatatcaa ggagctgctg gagcccatgc tggcagtggg actaagccct gccctcactg
     1741 cagtgctcta cgacctgagc cgtcagattc cacagctaaa gaaggacatt caagatgggc
     1801 tactgaaaat gctgtccctg gtccttatgc acaaacccct tcgccaccca ggcatgccca
     1861 agggcctggc ccatcagctg gcctctcctg gcctcacgac cctccctgag gccagcgatg
     1921 tgggcagcat cactcttgcc ctccgaacgc ttggcagctt tgaatttgaa ggccactctc
     1981 tgacccaatt tgttcgccac tgtgcggatc attttctgaa cagtgagcac aaggagatcc
     2041 gcatggaggc tgcccgcacc tgctcccgcc tgctcacacc ctccatccac ctcatcagtg
     2101 gccatgctca tgtggttagc cagaccgcag tgcaagtggt ggcagatgtg cttagcaaac
     2161 tgctcgtagt tgggataaca gatcctgacc ctgacattcg ctactgtgtc ttggcgtccc
     2221 tggacgagcg ctttgatgca cacctggccc aggcggagaa cttgcaggcc ttgtttgtgg
     2281 ctctgaatga ccaggtgttt gagatccggg agctggccat ctgcactgtg ggccgactca
     2341 gtagcatgaa ccctgccttt gtcatgcctt tcctgcgcaa gatgctcatc cagattttga
     2401 cagagttgga gcacagtggg attggaagaa tcaaagagca gagtgcccgc atgctggggc
     2461 acctggtctc caatgccccc cgactcatcc gcccctacat ggagcctatt ctgaaggcat
     2521 taattttgaa actgaaagat ccagaccctg atccaaaccc aggtgtgatc aataatgtcc
     2581 tggcaacaat aggagaattg gcacaggtta gtggcctgga aatgaggaaa tgggttgatg
     2641 aactttttat tatcatcatg gacatgctcc aggattcctc tttgttggcc aaaaggcagg
     2701 tggctctgtg gaccctggga cagttggtgg ccagcactgg ctatgtagta gagccctaca
     2761 ggaagtaccc tactttgctt gaggtgctac tgaattttct gaagactgag cagaaccagg
     2821 gtacacgcag agaggccatc cgtgtgttag ggcttttagg ggctttggat ccttacaagc
     2881 acaaagtgaa cattggcatg atagaccagt cccgggatgc ctctgctgtc agcctgtcag
     2941 aatccaagtc aagtcaggat tcctctgact atagcactag tgaaatgctg gtcaacatgg
     3001 gaaacttgcc tctggatgag ttctacccag ctgtgtccat ggtggccctg atgcggatct
     3061 tccgagacca gtcactctct catcatcaca ccatggttgt ccaggccatc accttcatct
     3121 tcaagtccct gggactcaaa tgtgtgcagt tcctgcccca ggtcatgccc acgttcctta
     3181 atgtcattcg agtctgtgat ggggccatcc gggaattttt gttccagcag ctgggaatgt
     3241 tggtgtcctt tgtgaagagc cacatcagac cttatatgga tgaaatagtc accctcatga
     3301 gagaattctg ggtcatgaac acctcaattc agagcacgat cattcttctc attgagcaaa
     3361 ttgtggtagc tcttgggggt gaatttaagc tctacctgcc ccagctgatc ccacacatgc
     3421 tgcgtgtctt catgcatgac aacagcccag gccgcattgt ctctatcaag ttactggctg
     3481 caatccagct gtttggcgcc aacctggatg actacctgca tttactgctg cctcctattg
     3541 ttaagttgtt tgatgcccct gaagctccac tgccatctcg aaaggcagcg ctagagactg
     3601 tggaccgcct gacggagtcc ctggatttca ctgactatgc ctcccggatc attcacccta
     3661 ttgttcgaac actggaccag agcccagaac tgcgctccac agccatggac acgctgtctt
     3721 cacttgtttt tcagctgggg aagaagtacc aaattttcat tccaatggtg aataaagttc
     3781 tggtgcgaca ccgaatcaat catcagcgct atgatgtgct catctgcaga attgtcaagg
     3841 gatacacact tgctgatgaa gaggaggatc ctttgattta ccagcatcgg atgcttagga
     3901 gtggccaagg ggatgcattg gctagtggac cagtggaaac aggacccatg aagaaactgc
     3961 acgtcagcac catcaacctc caaaaggcct ggggcgctgc caggagggtc tccaaagatg
     4021 actggctgga atggctgaga cggctgagcc tggagctgct gaaggactca tcatcgccct
     4081 ccctgcgctc ctgctgggcc ctggcacagg cctacaaccc gatggccagg gatctcttca
     4141 atgctgcatt tgtgtcctgc tggtctgaac tgaatgaaga tcaacaggat gagctcatca
     4201 gaagcatcga gttggccctc acctcacaag acatcgctga agtcacacag accctcttaa
     4261 acttggctga attcatggaa cacagtgaca agggccccct gccactgaga gatgacaatg
     4321 gcattgttct gctgggtgag agagctgcca agtgccgagc atatgccaaa gcactacact
     4381 acaaagaact ggagttccag aaaggcccca cccctgccat tctagaatct ctcatcagca
     4441 ttaataataa gctacagcag ccggaggcag cggccggagt gttagaatat gccatgaaac
     4501 actttggaga gctggagatc caggctacct ggtatgagaa actgcacgag tgggaggatg
     4561 cccttgtggc ctatgacaag aaaatggaca ccaacaagga cgacccagag ctgatgctgg
     4621 gccgcatgcg ctgcctcgag gccttggggg aatggggtca actccaccag cagtgctgtg
     4681 aaaagtggac cctggttaat gatgagaccc aagccaagat ggcccggatg gctgctgcag
     4741 ctgcatgggg tttaggtcag tgggacagca tggaagaata cacctgtatg atccctcggg
     4801 acacccatga tggggcattt tatagagctg tgctggcact gcatcaggac ctcttctcct
     4861 tggcacaaca gtgcattgac aaggccaggg acctgctgga tgctgaatta actgcgatgg
     4921 caggagagag ttacagtcgg gcatatgggg ccatggtttc ttgccacatg ctgtccgagc
     4981 tggaggaggt tatccagtac aaacttgtcc ccgagcgacg agagatcatc cgccagatct
     5041 ggtgggagag actgcagggc tgccagcgta tcgtagagga ctggcagaaa atccttatgg
     5101 tgcggtccct tgtggtcagc cctcatgaag acatgagaac ctggctcaag tatgcaagcc
     5161 tgtgcggcaa gagtggcagg ctggctcttg ctcataaaac tttagtgttg ctcctgggag
     5221 ttgatccgtc tcggcaactt gaccatcctc tgccaacagt tcaccctcag gtgacctatg
     5281 cctacatgaa aaacatgtgg aagagtgccc gcaagatcga tgccttccag cacatgcagc
     5341 attttgtcca gaccatgcag caacaggccc agcatgccat cgctactgag gaccagcagc
     5401 ataagcagga actgcacaag ctcatggccc gatgcttcct gaaacttgga gagtggcagc
     5461 tgaatctaca gggcatcaat gagagcacaa tccccaaagt gctgcagtac tacagcgccg
     5521 ccacagagca cgaccgcagc tggtacaagg cctggcatgc gtgggcagtg atgaacttcg
     5581 aagctgtgct acactacaaa catcagaacc aagcccgcga tgagaagaag aaactgcgtc
     5641 atgccagcgg ggccaacatc accaacgcca ccactgccgc caccacggcc gccactgcca
     5701 ccaccactgc cagcaccgag ggcagcaaca gtgagagtga ggccgagagc accgagaaca
     5761 gccccacccc atcgccgctg cagaagaagg tcactgagga tctgtccaaa accctcctga
     5821 tgtacacggt gcctgccgtc cagggcttct tccgttccat ctccttgtca cgaggcaaca
     5881 acctccagga tacactcaga gttctcacct tatggtttga ttatggtcac tggccagatg
     5941 tcaatgaggc cttagtggag ggggtgaaag ccatccagat tgatacctgg ctacaggtta
     6001 tacctcagct cattgcaaga attgatacgc ccagaccctt ggtgggacgt ctcattcacc
     6061 agcttctcac agacattggt cggtaccacc cccaggccct catctaccca ctgacagtgg
     6121 cttctaagtc taccacgaca gcccggcaca atgcagccaa caagattctg aagaacatgt
     6181 gtgagcacag caacaccctg gtccagcagg ccatgatggt gagcgaggag ctgatccgag
     6241 tggccatcct ctggcatgag atgtggcatg aaggcctgga agaggcatct cgtttgtact
     6301 ttggggaaag gaacgtgaaa ggcatgtttg aggtgctgga gcccttgcat gctatgatgg
     6361 aacggggccc ccagactctg aaggaaacat cctttaatca ggcctatggt cgagatttaa
     6421 tggaggccca agagtggtgc aggaagtaca tgaaatcagg gaatgtcaag gacctcaccc
     6481 aagcctggga cctctattat catgtgttcc gacgaatctc aaagcagctg cctcagctca
     6541 catccttaga gctgcaatat gtttccccaa aacttctgat gtgccgggac cttgaattgg
     6601 ctgtgccagg aacatatgac cccaaccagc caatcattcg cattcagtcc atagcaccgt
     6661 ctttgcaagt catcacatcc aagcagaggc cccggaaatt gacacttatg ggcagcaacg
     6721 gacatgagtt tgttttcctt ctaaaaggcc atgaagatct gcgccaggat gagcgtgtga
     6781 tgcagctctt cggcctggtt aacaccctcc tggccaatga cccaacatct cttcggaaaa
     6841 acctcagcat ccagagatac gctgtcatcc ctttatcgac caactcgggc ctcattggct
     6901 gggttcccca ctgtgacaca ctgcacgccc tcatccggga ctacagggag aagaagaaga
     6961 tccttctcaa catcgagcat cgcatcatgt tgcggatggc tccggactat gaccacttga
     7021 ctctgatgca gaaggtggag gtgtttgagc atgccgtcaa taatacagct ggggacgacc
     7081 tggccaagct gctatggctg aaaagcccca gctccgaggt gtggtttgac cgaagaacca
     7141 attatacccg ttctttagcg gtcatgtcaa tggttgggta tattttaggc ctgggagata
     7201 gacacccatc caacctgatg ctggaccgtc tgagtgggaa gatcctgcac attgactttg
     7261 gggactgctt tgaggttgct atgacccgag agaagtttcc agagaagatt ccatttagac
     7321 taacaagaat gttgaccaat gctatggagg ttacaggcct ggatggcaac tacagaatca
     7381 catgccacac agtgatggag gtgctgcgag agcacaagga cagtgtcatg gccgtgctgg
     7441 aagcctttgt ctatgacccc ttgctgaact ggaggctgat ggacacaaat accaaaggca
     7501 acaagcgatc ccgaacgagg acggattcct actctgctgg ccagtcagtc gaaattttgg
     7561 acggtgtgga acttggagag ccagcccata agaaaacggg gaccacagtg ccagaatcta
     7621 ttcattcttt cattggagac ggtttggtga aaccagaggc cctaaataag aaagctatcc
     7681 agattattaa cagggttcga gataagctca ctggtcggga cttctctcat gatgacactt
     7741 tggatgttcc aacgcaagtt gagctgctca tcaaacaagc gacatcccat gaaaacctct
     7801 gccagtgcta tattggctgg tgccctttct ggtcaacttt cttgtacaaa gttggcatta
     7861 taggaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     7921 tcattatttg catgcaagct tagatcctct agaccagcca ggacagaaat gcctcgactt
     7981 cgctgctacc caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc
     8041 agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca
     8101 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     8161 cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc
     8221 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca
     8281 gtcgccctaa aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc
     8341 aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac
     8401 atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg
     8461 ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg
     8521 aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg
     8581 tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat
     8641 ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg
     8701 ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg
     8761 aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc
     8821 tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc
     8881 gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg
     8941 caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc
     9001 ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact
     9061 acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa
     9121 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     9181 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     9241 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     9301 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     9361 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     9421 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     9481 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     9541 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc
     9601 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     9661 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     9721 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     9781 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     9841 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     9901 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     9961 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
    10021 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
    10081 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
    10141 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
    10201 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
    10261 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
    10321 ggcagttccc tactctcgcg
//
