LOCUS       sequence-194179-        6193 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresse Myc tagged metap2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6193)
  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 6193)
  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-metap2
FEATURES             Location/Qualifiers
     source          1..6193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      950..959
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             959..988
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      2480..2489
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2493..2627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2530..2549)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2584..2603
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2696..3053
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2696..2887
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2719..2739)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2904..3039
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3073..3090)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3073..3089)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3082..3104)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3302..3757
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3389..3408)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3599..3620
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3774..3793)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3893..3915
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3953..3971)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4039..4143
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4144..4212,4213..5004)
                     /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(4362..4381)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5175..5763
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5664..5683
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5917..5934
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6051..6072
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6087..6092,6093..6110,6111..6117)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6125..6141
                     /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     6130..6152
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6149..6165
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6149..6165
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgattgaatt cccgaaccga cagtcggtct cttcaccaag gccattcgcg ccaccatgga
      961 gcaaaagctc atttctgagg aagatctcaa tggtggtggt ggtgggtcga cgatggcggg
     1021 tgtggaggag gtagcggcct ccgggagcca cctgaatggc gacctggatc cagacgacag
     1081 ggaagaagga gctgcctcta cggctgagga agcagccaag aaaaaaagac gaaagaagaa
     1141 gaagagcaaa gggccttctg caggggaaca ggaacctgat aaagaatcag gagcctcagt
     1201 ggatgaagta gcaagacagt tggaaagatc agcattggaa gataaagaaa gagatgaaga
     1261 tgatgaagat ggagatggcg atggagatgg agcaactgga aagaagaaga aaaagaagaa
     1321 gaagaagaga ggaccaaaag ttcaaacaga ccctccctca gttccaatat gtgacctgta
     1381 tcctaatggt gtatttccca aaggacaaga atgcgaatac ccacccacac aagatgggcg
     1441 aacagctgct tggagaacta caagtgaaga aaagaaagca ttagatcagg caagtgaaga
     1501 gatttggaat gattttcgag aagctgcaga agcacatcga caagttagaa aatacgtaat
     1561 gagctggatc aagcctggga tgacaatgat agaaatctgt gaaaagttgg aagactgttc
     1621 acgcaagtta ataaaagaga atggattaaa tgcaggcctg gcatttccta ctggatgttc
     1681 tctcaataat tgtgctgccc attatactcc caatgccggt gacacaacag tattacagta
     1741 tgatgacatc tgtaaaatag actttggaac acatataagt ggtaggatta ttgactgtgc
     1801 ttttactgtc acttttaatc ccaaatatga tacgttatta aaagctgtaa aagatgctac
     1861 taacactgga ataaagtgtg ctggaattga tgttcgtctg tgtgatgttg gtgaggccat
     1921 ccaagaagtt atggagtcct atgaagttga aatagatggg aagacatatc aagtgaaacc
     1981 aatccgtaat ctaaatggac attcaattgg gcaatataga atacatgctg gaaaaacagt
     2041 gccgattgtg aaaggagggg aggcaacaag aatggaggaa ggagaagtat atgcaattga
     2101 aacctttggt agtacaggaa aaggtgttgt tcatgatgat atggaatgtt cacattacat
     2161 gaaaaatttt gatgttggac atgtgccaat aaggcttcca agaacaaaac acttgttaaa
     2221 tgtcatcaat gaaaactttg gaacccttgc cttctgccgc agatggctgg atcgcttggg
     2281 agaaagtaaa tacttgatgg ctctgaagaa tctgtgtgac ttgggcattg tagatccata
     2341 tccaccatta tgtgacatta aaggatcata tacagcgcaa tttgaacata ccatcctgtt
     2401 gcgtccaaca tgtaaagaag ttgtcagcag aggagatgac tattaagcgg ccgctaagta
     2461 agtaaggatc cccagcttgg ccgccatggc ccaacttgtt tattgcagct tataatggtt
     2521 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2581 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcggg aattaattcg
     2641 gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag
     2701 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2761 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     2821 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2881 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2941 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     3001 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctgt
     3061 taacagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     3121 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     3181 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     3241 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     3301 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3361 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3421 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3481 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     3541 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3601 ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta
     3661 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3721 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     3781 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     3841 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     3901 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     3961 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     4021 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4081 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4141 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4201 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4261 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4321 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4381 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4441 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4501 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4561 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4621 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4681 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4741 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4801 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4861 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4921 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4981 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5041 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5101 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5161 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5221 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5281 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5341 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5401 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5461 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5521 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5581 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5641 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5701 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5761 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5821 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5881 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5941 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     6001 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     6061 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     6121 ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat tacgaattaa
     6181 ttcgagctcg ccc
//
