LOCUS       sequence-197110-        5977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Myc tagged SAMTOR in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5977)
  AUTHORS   Gu X, Orozco JM, Saxton RA, Condon KJ, Liu GY, Krawczyk PA, Scaria 
            SM, Harper JW, Gygi SP, Sabatini DM
  TITLE     SAMTOR is an S-adenosylmethionine sensor for the mTORC1 pathway.
  JOURNAL   Science. 2017 Nov 10;358(6364):813-818. doi: 
            10.1126/science.aao3265.
  PUBMED    29123071
REFERENCE   2  (bases 1 to 5977)
  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"; 
            journalName: "Science"; date: "2017-11-10- 10"; volume: "358"; 
            issue: "6364"; pages: "813-818"
COMMENT     Sequence Label: pRK5-Myc-SAMTOR
FEATURES             Location/Qualifiers
     source          1..5977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        144..523
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        524..727
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     678..698
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     724..748
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        958..976
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     958..975
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     989..1008
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1093..1102
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1102..1131
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      2407..2416
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2420..2554
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2457..2476)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2511..2530
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2623..2980
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2623..2814
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2646..2666)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2831..2966
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3000..3017)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3000..3016)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3009..3031)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3229..3684
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3316..3335)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3526..3547
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3701..3720)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3820..3842
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3880..3898)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3966..4070
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4071..4139,4140..4931)
                     /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(4289..4308)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5102..5690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5591..5610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5844..5861
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat
      121 tacgaattaa ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat
      181 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      241 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      301 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      361 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      421 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      481 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      541 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      601 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      661 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      721 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
      781 ccatagaaga caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg
      841 gattccccgt gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt
      901 ggcttcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
      961 taggtgacac tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag
     1021 gtccaactgc acctcggttc tatcgattga attcccgaac cgacagtcgg tctcttcacc
     1081 aaggccattc gcgccaccat ggagcaaaag ctcatttctg aggaagatct caatggtggt
     1141 ggtggtgggt cgacgatgga gccaggggcc ggcggccgaa atactgctcg tgcgcagagg
     1201 gccgggtccc cgaacactcc gccgccccgg gagcaggagc ggaaactgga gcaggagaag
     1261 ctctccggtg tggtgaagag cgtccaccgg cggctccgca agaagtaccg agaagtggga
     1321 gattttgata agatctggcg agaacactgt gaggatgaag aaactctttg tgaatatgcc
     1381 gttgcaatga aaaatttggc agataatcat tgggcaaaaa cttgtgaggg cgaaggtcgt
     1441 attgaatggt gttgtagtgt atgcagagaa tatttccaaa atggtgggaa gagaaaagca
     1501 cttgagaaag atgaaaaaag agctgtactt gccactaaga ccactccagc cttaaatatg
     1561 catgagtctt ctcaacttga aggtcattta accaacctca gctttacaaa ccctgaattt
     1621 ataactgagt tgctacaagc ctcaggaaaa atcagattac ttgatgttgg cagctgcttt
     1681 aacccatttc tgaagtttga agaatttcta actgttggca tagatattgt acctgctgta
     1741 gagagtgtct ataaatgtga tttcctgaac ttacagcttc agcaaccact ccagcttgca
     1801 caggatgcta tagatgcttt tttgaagcag ctgaaaaacc ctattgattc tcttcctgga
     1861 gagcttttcc atgtggttgt tttctctctc cttctttctt attttccatc tccttaccag
     1921 cgatggattt gctgcaagaa agcccatgaa ctgttagtgt taaatggttt attactaatc
     1981 atcacacctg attcctccca tcagaaccgt catgctatga tgatgaaaag ctggaagatt
     2041 gctatagagt ccctgggctt taaacgcttc aagtactcaa aattttcaca tatgcatctg
     2101 atggcattta ggaaaatctc tctaaaaacc acaagtgact tggttagtag gaactaccca
     2161 ggaatgttat atattcctca agatttcaac agtatagaag atgaggaata ttctaaccct
     2221 tcctgctatg ttcgatcaga tatagaagat gaacaactag catatggttt cacagaactc
     2281 cctgatgcgc catatgactc agattctgga gaaagtcaag ccagctctat tcctttctat
     2341 gagctagaag accccatatt acttttaagt taagcggccg ctaagtaagt aaggatcccc
     2401 agcttggccg ccatggccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     2461 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2521 caaactcatc aatgtatctt atcatgtctg gatcgggaat taattcggcg cagcaccatg
     2581 gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca
     2641 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2701 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2761 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2821 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2881 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     2941 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctgttaa cagcttggca
     3001 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     3061 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     3121 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt
     3181 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     3241 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3301 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3361 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3421 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     3481 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3541 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc
     3601 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3661 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     3721 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3781 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3841 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     3901 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     3961 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4021 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4081 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4141 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4201 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4261 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4321 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4381 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4441 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4501 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4561 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4621 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4681 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4741 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4801 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4861 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4921 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4981 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5041 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5101 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5161 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5221 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5281 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5341 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5401 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5461 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5521 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5581 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5641 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5701 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5761 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     5821 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     5881 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     5941 ttcccgactg gaaagcgggc agtgagcgca acgcaat
//
