LOCUS       sequence-474947-        8117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of human WDR24-SEH1L ""single chain"" in the 
            GATOR2 complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8117)
  AUTHORS   Valenstein ML, Wranik M, Lalgudi PV, Linde-Garelli KY, Choi Y, 
            Chivukula RR, Sabatini DM, Rogala KB
  TITLE     Structural basis for the dynamic regulation of mTORC1 by amino 
            acids.
  JOURNAL   Nature. 2025 Aug 20. doi: 10.1038/s41586-025-09428-7.
  PUBMED    40836086
REFERENCE   2  (bases 1 to 8117)
  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"; journalName: "Nature. 
            2025 Aug 20. doi: 10.1038/s41586-025-09428-7."
COMMENT     Sequence Label: sc-WDR24-SEH1L
FEATURES             Location/Qualifiers
     source          1..8117
                     /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"
     regulatory      925..934
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4417..4426
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4430..4564
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4467..4486)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4521..4540
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4633..4990
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4633..4824
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4656..4676)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4841..4976
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5010..5027)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5010..5026)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5019..5041)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5239..5694
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5326..5345)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5536..5557
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5711..5730)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5830..5852
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5890..5908)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5976..6080
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6081..6149,6150..6941)
                     /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(6299..6318)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7112..7700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7601..7620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7854..7871
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7988..8009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8024..8029,8030..8047,8048..8054)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8062..8078
                     /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     8067..8089
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8086..8102
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8086..8102
                     /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 tatcgattga attcgccacc atggagaaga tgtcccgtgt gaccacagcc
      961 ctgggtggca gcgtgctgac aggccgcacc atgcactgcc acctggatgc tcccgccaat
     1021 gccatcagtg tgtgccgcga cgcagcccag gtggtcgtgg caggccgtag catcttcaag
     1081 atctatgcca tcgaggagga acagttcgtg gaaaagctga acctgcgtgt ggggcgcaag
     1141 ccttcgctta acctgagctg tgctgacgtg gtctggcacc agatggatga gaacctgctg
     1201 gccacagcag ccaccaatgg cgtggtggtc acgtggaacc tgggccggcc atcccgcaac
     1261 aagcaggacc agctgttcac agaacacaag cgcacggtaa acaaagtctg cttccacccc
     1321 accgaagccc acgtgctgct cagtggctcc caggatggct tcatgaagtg ctttgacctc
     1381 cgcagaaagg actctgtcag caccttctcg ggccagtcgg agagcgtgcg ggacgtgcag
     1441 ttcagtatcc gggactactt caccttcgcc tccacctttg agaacggcaa tgtgcagctc
     1501 tgggacatcc ggcgtcccga ccggtgcgag aggatgttca cagcccacaa cggacccgtc
     1561 ttctgctgcg actggcaccc cgaggacagg ggctggttgg ccactggagg gcgcgacaag
     1621 atggtgaagg tctgggacat gaccacgcac cgtgccaagg agatgcactg tgtgcagacc
     1681 atcgcctcgg tggcccgtgt gaagtggcgg ccagagtgcc gccaccacct ggccacgtgc
     1741 tccatgatgg tggaccacaa catctatgtt tgggacgtgc gccggccctt cgtgccagct
     1801 gccatgtttg aggaacaccg agacgtcacc acgggaattg cctggcgcca cccccacgac
     1861 ccctccttcc tgctgtctgg ctccaaggac agctcgctgt gccagcacct gttccgcgac
     1921 gccagccagc ccgtcgagcg cgccaaccct gagggcctct gctacggcct cttcggggac
     1981 ctggccttcg ccgccaagga gagcctcgtg gctgccgagt cggggcgcaa gccctacact
     2041 ggcgaccggc gccaccccat cttctttaag cgcaagctgg accctgccga gcccttcgca
     2101 ggcctcgcct ccagtgccct cagtgtcttt gagacggagc caatgtttgt ggctcgcagc
     2161 atcgcggcgg accacaagga tctcatccac gatgtctctt tcgacttcca cgggcggcgg
     2221 atggcaacct gctccagcga tcagagcgtt aaggtctggg ataaaagtga aagtggtgat
     2281 tggcattgta ctgctagctg gaagacacat agtggatctg tatggcgtgt gacatgggcc
     2341 catcctgaat ttgggcaggt tttggcttcc tgttcttttg accgaacagc tgctgtatgg
     2401 gaagaaatag taggagaatc aaatgataaa ctgcgaggac agagccactg ggttaaaagg
     2461 acaactctgg tggatagcag aacatctgtt actgatgtga agtttgctcc caagcacatg
     2521 ggtcttatgt tagcaacctg ttccgcagat ggtatagtaa gaatctatga ggcaccagat
     2581 gttatgaatc tcagccagtg gtctttgcag catgagatct catgtaagct aagctgtagt
     2641 tgtatttctt ggaacccttc aagctctcgt gctcattccc ccatgatcgc cgtaggaagt
     2701 gatgacagta gccccaacgc aatggccaag gttcagattt ttgaatataa tgaaaacacc
     2761 aggaaatatg caaaagctga aactcttatg acagtcactg atcctgttca tgatattgca
     2821 ttcgctccaa atttgggaag atctttccat attctagcaa tagcgaccaa agatgtgaga
     2881 atttttacat taaagcctgt gaggaaagaa ctgacttcct ctggtgggcc aacaaagttt
     2941 gaaatccata tagtggctca gttcgataat cataattctc aggtctggcg agtgagttgg
     3001 aatataacag gaacggtgct agcatcttca ggagatgatg ggtgtgtaag attgtggaaa
     3061 gctaattata tggacaattg gaagtgtact ggtattttga aaggtaatgg gagcccagtc
     3121 aatgggagtt ctcagcaggg aacctcaaat ccttccctag gttcaactat tccaagtctt
     3181 cagaattcat taaatggatc ttctgctggc agaaagcaca gcggtggcgg cggcatgcgc
     3241 tggtttgtgg acacagctga gcgttatgcg ctggctggcc ggccactggc cgagctctgt
     3301 gaccacaacg caaaggtggc tcgagagctt ggccgcaacc aggtggcgca aacgtggacc
     3361 atgctgcgga tcatctactg cagccctggc ctagtgccca ctgcaaacct caaccacagt
     3421 gtgggcaagg gtggctcctg tggcctcccg ctcatgaaca gtttcaacct gaaggatatg
     3481 gccccagggt tgggcagtga gacgcggctg gaccgcagca aaggagatgc gcggagcgac
     3541 acagttctgc tcgactcctc ggccacactc atcaccaatg aggataacga ggaaaccgag
     3601 ggcagcgacg tacctgccga ctacctgctg ggtgacgtgg aaggtgagga ggacgagctg
     3661 tacctgctgg atccggaaca cgcgcacccc gaggaccctg agtgcgtgct gccgcaggag
     3721 gcctttccgc tgcgccacga gatcgtggac acgcctcccg ggcccgagca cctgcaggac
     3781 aaggccgact ccccgcacgt gagcggcagc gaggcggatg tggcctccct ggcccccgtg
     3841 gactcctcct tctcgctcct gtctgtctca cacgcgctct acgacagccg cctgccgccc
     3901 gacttcttcg gcgtgctggt gcgcgacatg ctgcacttct acgctgagca gggcgacgtg
     3961 cagatggctg tgtctgtgct catcgtcctg ggtgaacggg tgcgcaagga catcgacgag
     4021 cagacccagg agcactggta cacttcctac atcgacctgc tgcagcgctt ccgcctctgg
     4081 aacgtgtcca acgaggtggt caagctgagc accagccgcg ccgtcagctg cctcaaccag
     4141 gcctccacca ccctgcacgt caactgcagc cactgcaagc ggcccatgag cagccggggc
     4201 tgggtctgcg acaggtgcca ccgctgcgcc agcatgtgtg ccgtctgcca ccacgtagtc
     4261 aagggtctct tcgtgtggtg ccagggctgc agccacggcg gccacctgca gcacatcatg
     4321 aagtggctgg aaggcagctc ccactgtccc gcaggctgcg gccacctctg cgagtactcc
     4381 tgagcggccg ctaagtaagt aaggatcccc agcttggccg ccatggccca acttgtttat
     4441 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4501 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4561 gatcgggaat taattcggcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg
     4621 gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     4681 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4741 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4801 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4861 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4921 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4981 cttttgcaaa aagctgttaa cagcttggca ctggccgtcg ttttacaacg tcgtgactgg
     5041 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     5101 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     5161 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     5221 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     5281 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     5341 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     5401 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     5461 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5521 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     5581 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     5641 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     5701 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     5761 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5821 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     5881 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     5941 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6001 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6061 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6121 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6181 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6241 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6301 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6361 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6421 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6481 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6541 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6601 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6661 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6721 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6781 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6841 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6901 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6961 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7021 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7081 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7141 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7201 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     7261 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7321 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7381 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7441 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7501 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7561 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7621 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7681 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7741 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     7801 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     7861 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     7921 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     7981 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     8041 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     8101 acatgattac gaattaa
//
