LOCUS       sequence-474946-        8413 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of human MIOS-SEH1L ""single chain"" in the 
            GATOR2 complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8413)
  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 8413)
  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-MIOS-SEH1L
FEATURES             Location/Qualifiers
     source          1..8413
                     /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             4666..4692
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      4713..4722
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4726..4860
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4763..4782)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4817..4836
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4929..5286
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4929..5120
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4952..4972)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5137..5272
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5306..5323)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5306..5322)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5315..5337)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5535..5990
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5622..5641)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5832..5853
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6007..6026)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6126..6148
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6186..6204)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6272..6376
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6377..6445,6446..7237)
                     /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(6595..6614)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7408..7996
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7897..7916
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8150..8167
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8284..8305
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8320..8325,8326..8343,8344..8350)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8358..8374
                     /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     8363..8385
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8382..8398
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8382..8398
                     /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 attcgtcgac gccaccatgt cgggcaccaa accggatatt
      961 ctgtgggcac cgcatcatgt tgaccgcttc gttgtctgtg actcggaact gtccctgtac
     1021 catgtcgaaa gtaccgtgaa cagcgaactg aaagcaggca gcctgcgtct gtcagaagac
     1081 tcggcggcca ccctgctgag cattaactct gatacgccgt atatgaaatg tgtcgcgtgg
     1141 tatctgaact acgatccgga atgcctgctg gccgttggcc aggcaaatgg tcgcgtggtt
     1201 ctgaccagcc tgggccaaga ccacaactca aaattcaaag atctgatcgg taaagaattt
     1261 gtgccgaaac atgcccgtca gtgtaacacc ctggcatgga atccgctgga ttctaactgg
     1321 ctggcagctg gcctggacaa acaccgcgcg gatttttctg tgctgatttg ggacatctgc
     1381 agtaaatata ccccggatat tgttccgatg gaaaaagtca aactgtccgc tggcgaaacc
     1441 gaaaccacgc tgctggttac gaaaccgctg tacgaactgg gtcagaatga cgcctgtctg
     1501 tcactgtgct ggctgccgcg tgatcaaaaa ctgctgctgg ctggtatgca ccgtaatctg
     1561 gcgatttttg atctgcgcaa cacctcgcag aaaatgttcg tgaataccaa agcggttcaa
     1621 ggcgtgacgg ttgacccgta ttttcatgat cgtgtcgcta gcttctacga gggtcaggtg
     1681 gcgatctggg atctgcgcaa atttgaaaaa ccagtgctga ccctgacgga acaaccgaaa
     1741 ccgctgacca aagtcgcatg gtgtccgacc cgtacgggcc tgctggcaac cctgacgcgt
     1801 gactctaata ttatccgcct gtatgatatg cagcacaccc cgacgccgat tggtgacgaa
     1861 accgaaccga cgattatcga acgcagtgtg cagccgtgcg ataactacat tgcttcgttc
     1921 gcgtggcatc cgaccagcca aaaccgtatg atcgtcgtga ccccgaatcg cacgatgagc
     1981 gattttaccg ttttcgaacg tatttcactg gcttggtcgc cgatcacgag cctgatgtgg
     2041 gcgtgtggtc gccacctgta tgaatgcacc gaagaaatgt ttgtggctcg cagcatcgcg
     2101 gcggaccaca aggatctcat ccacgatgtc tctttcgact tccacgggcg gcggatggca
     2161 acctgctcca gcgatcagag cgttaaggtc tgggataaaa gtgaaagtgg tgattggcat
     2221 tgtactgcta gctggaagac acatagtgga tctgtatggc gtgtgacatg ggcccatcct
     2281 gaatttgggc aggttttggc ttcctgttct tttgaccgaa cagctgctgt atgggaagaa
     2341 atagtaggag aatcaaatga taaactgcga ggacagagcc actgggttaa aaggacaact
     2401 ctggtggata gcagaacatc tgttactgat gtgaagtttg ctcccaagca catgggtctt
     2461 atgttagcaa cctgttccgc agatggtata gtaagaatct atgaggcacc agatgttatg
     2521 aatctcagcc agtggtcttt gcagcatgag atctcatgta agctaagctg tagttgtatt
     2581 tcttggaacc cttcaagctc tcgtgctcat tcccccatga tcgccgtagg aagtgatgac
     2641 agtagcccca acgcaatggc caaggttcag atttttgaat ataatgaaaa caccaggaaa
     2701 tatgcaaaag ctgaaactct tatgacagtc actgatcctg ttcatgatat tgcattcgct
     2761 ccaaatttgg gaagatcttt ccatattcta gcaatagcga ccaaagatgt gagaattttt
     2821 acattaaagc ctgtgaggaa agaactgact tcctctggtg ggccaacaaa gtttgaaatc
     2881 catatagtgg ctcagttcga taatcataat tctcaggtct ggcgagtgag ttggaatata
     2941 acaggaacgg tgctagcatc ttcaggagat gatgggtgtg taagattgtg gaaagctaat
     3001 tatatggaca attggaagtg tactggtatt ttgaaaggta atgggagccc agtcaatggg
     3061 agttctcagc agggaacctc aaatccttcc ctaggttcaa ctattccaag tcttcagaat
     3121 tcattaaatg gatcttctgc tggcagaaag cacagcgaaa acgacaattc gctggaaaaa
     3181 gatattgcca cgaaaatgcg tctgcgcgca ctgagccgtt acggcctgga caccgaacaa
     3241 gtgtggcgca atcacatcct ggccggtaac gaagatccgc aactgaaatc cctgtggtat
     3301 accctgcatt ttatgaaaca gtacacggaa gatatggacc aaaaatcgcc gggcaataaa
     3361 ggtagcctgg tttatgcggg cattaaaagc atcgtgaaaa gctctctggg tatggttgaa
     3421 agttcccgtc ataactggtc tggcctggat aaacagagtg acattcaaaa cctgaatgaa
     3481 gaacgcatcc tggccctgca gctgtgtggc tggattaaga aaggcaccga tgtggatgtg
     3541 ggcccgtttc tgaacagcct ggtgcaagaa ggtgaatggg aacgcgcggc ggcagttgcg
     3601 ctgttcaacc tggatattcg tcgcgccatt cagatcctga atgaaggcgc atcatcggaa
     3661 aaaggtgatc tgaacctgaa tgttgtcgct atggcgctga gtggttatac cgatgagaaa
     3721 aacagcctgt ggcgtgaaat gtgtagtacg ctgcgcctgc agctgaacaa tccgtacctg
     3781 tgcgtgatgt ttgctttcct gacctctgaa acgggcagtt atgatggtgt cctgtacgaa
     3841 aataaagttg cggtccgtga ccgcgtggcc tttgcatgta aattcctgtc agatacccag
     3901 ctgaaccgtt atatcgaaaa actgacgaac gaaatgaaag aagccggcaa cctggaaggt
     3961 atcctgctga ccggcctgac gaaagatggt gttgacctga tggaaagcta tgtcgatcgc
     4021 accggcgacg tgcagacggc atcctactgc atgctgcaag gttcaccgct ggatgttctg
     4081 aaagacgaac gtgtccagta ttggattgaa aactaccgca atctgctgga tgcttggcgt
     4141 ttttggcaca aacgcgcgga atttgacatc catcgttcta aactggatcc gagctctaaa
     4201 ccgctggccc aggtgtttgt tagttgtaac ttctgcggca aatcaatttc gtatagctgc
     4261 tctgcagtgc cgcatcaggg tcgcggtttt agccaatacg gcgttagtgg ttccccgacc
     4321 aaatccaaag tcacgtcatg tccgggttgc cgtaaaccgc tgccgcgttg tgcactgtgc
     4381 ctgatcaata tgggtacccc ggtgtcttcc tgtccgggcg gtacgaaatc ggatgaaaaa
     4441 gttgacctga gcaaagataa aaaactggcc cagtttaaca attggttcac ctggtgtcac
     4501 aactgccgtc atggcggtca cgcaggtcat atgctgtcct ggtttcgcga tcatgctgaa
     4561 tgcccggtgt cagcgtgcac ctgtaaatgc atgcaactgg atacgacggg taatctggtc
     4621 ccggcggaaa ccgttcaacc gtaagcggcc gcaggaggtg gaggttaccc ctatgatgtg
     4681 cctgactacg catgataagg atccccagct tggccgccat ggcccaactt gtttattgca
     4741 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4801 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     4861 gggaattaat tcggcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
     4921 ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     4981 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5041 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5101 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     5161 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5221 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     5281 tgcaaaaagc tgttaacagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     5341 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     5401 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     5461 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     5521 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5581 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     5641 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     5701 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     5761 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5821 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta
     5881 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5941 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     6001 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     6061 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     6121 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     6181 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     6241 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     6301 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     6361 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     6421 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     6481 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     6541 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     6601 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     6661 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     6721 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6781 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6841 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6901 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     6961 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     7021 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     7081 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     7141 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     7201 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     7261 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     7321 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     7381 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     7441 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     7501 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     7561 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     7621 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7681 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     7741 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7801 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7861 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7921 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7981 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     8041 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     8101 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     8161 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     8221 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     8281 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     8341 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacat
     8401 gattacgaat taa
//
