LOCUS       sequence-197111-        5936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG tagged SAMTOR in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5936)
  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 5936)
  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-FLAG-SAMTOR
FEATURES             Location/Qualifiers
     source          1..5936
                     /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"
     CDS             1067..1090
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2366..2375
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2379..2513
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2416..2435)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2470..2489
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2582..2939
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2582..2773
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2605..2625)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2790..2925
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2959..2976)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2959..2975)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2968..2990)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3188..3643
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3275..3294)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3485..3506
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3660..3679)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3779..3801
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3839..3857)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3925..4029
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4030..4098,4099..4890)
                     /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(4248..4267)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5061..5649
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5550..5569
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5803..5820
                     /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 attcaccatg gaaatggact acaaggatga
     1081 cgatgacaag ggtggtggtg gtggtgcgtc gacgatggag ccaggggccg gcggccgaaa
     1141 tactgctcgt gcgcagaggg ccgggtcccc gaacactccg ccgccccggg agcaggagcg
     1201 gaaactggag caggagaagc tctccggtgt ggtgaagagc gtccaccggc ggctccgcaa
     1261 gaagtaccga gaagtgggag attttgataa gatctggcga gaacactgtg aggatgaaga
     1321 aactctttgt gaatatgccg ttgcaatgaa aaatttggca gataatcatt gggcaaaaac
     1381 ttgtgagggc gaaggtcgta ttgaatggtg ttgtagtgta tgcagagaat atttccaaaa
     1441 tggtgggaag agaaaagcac ttgagaaaga tgaaaaaaga gctgtacttg ccactaagac
     1501 cactccagcc ttaaatatgc atgagtcttc tcaacttgaa ggtcatttaa ccaacctcag
     1561 ctttacaaac cctgaattta taactgagtt gctacaagcc tcaggaaaaa tcagattact
     1621 tgatgttggc agctgcttta acccatttct gaagtttgaa gaatttctaa ctgttggcat
     1681 agatattgta cctgctgtag agagtgtcta taaatgtgat ttcctgaact tacagcttca
     1741 gcaaccactc cagcttgcac aggatgctat agatgctttt ttgaagcagc tgaaaaaccc
     1801 tattgattct cttcctggag agcttttcca tgtggttgtt ttctctctcc ttctttctta
     1861 ttttccatct ccttaccagc gatggatttg ctgcaagaaa gcccatgaac tgttagtgtt
     1921 aaatggttta ttactaatca tcacacctga ttcctcccat cagaaccgtc atgctatgat
     1981 gatgaaaagc tggaagattg ctatagagtc cctgggcttt aaacgcttca agtactcaaa
     2041 attttcacat atgcatctga tggcatttag gaaaatctct ctaaaaacca caagtgactt
     2101 ggttagtagg aactacccag gaatgttata tattcctcaa gatttcaaca gtatagaaga
     2161 tgaggaatat tctaaccctt cctgctatgt tcgatcagat atagaagatg aacaactagc
     2221 atatggtttc acagaactcc ctgatgcgcc atatgactca gattctggag aaagtcaagc
     2281 cagctctatt cctttctatg agctagaaga ccccatatta cttttaagtt aagcggccgc
     2341 taagtaagta aggatcccca gcttggccgc catggcccaa cttgtttatt gcagcttata
     2401 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2461 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcgggaatt
     2521 aattcggcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
     2581 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2641 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2701 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2761 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2821 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     2881 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     2941 agctgttaac agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     3001 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     3061 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     3121 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     3181 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3241 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3301 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3361 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     3421 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3481 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt
     3541 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3601 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     3661 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3721 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3781 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     3841 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     3901 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3961 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4021 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4081 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4141 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4201 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4261 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4321 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4381 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4441 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4501 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4561 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4621 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4681 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4741 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4801 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4861 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4921 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4981 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5041 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5101 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5161 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5221 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5281 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5341 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5401 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5461 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5521 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5581 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5641 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5701 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5761 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5821 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     5881 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaat
//
