LOCUS       sequence-154102-        5150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses a constitutively active mutant of 4E-BP1 under the CAG 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5150)
  AUTHORS   Hartman NW, Lin TV, Zhang L, Paquelet GE, Feliciano DM, Bordey A
  TITLE     mTORC1 targets the translational repressor 4E-BP2, but not S6 kinase
            1/2, to regulate neural stem cell self-renewal in vivo.
  JOURNAL   Cell Rep. 2013 Oct 31;5(2):433-44. doi: 
            10.1016/j.celrep.2013.09.017. Epub 2013 Oct 17.
  PUBMED    24139800
REFERENCE   2  (bases 1 to 5150)
  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: 
            "10.1016/j.celrep.2013.09.017"; journalName: "Cell Rep"; date: 
            "2013-10-31- 31"; volume: "5"; issue: "2"; pages: "433-44"
COMMENT     Sequence Label: pCAGGS-4EBP1 F113A
FEATURES             Location/Qualifiers
     source          1..5150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..498
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        500..777
                     /label=chicken beta-actin promoter
     intron          778..1794
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1718..1740
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1802..1821
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(2286..2305)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2351..2406
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2405..2424)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2767..2783)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2767..2783)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2780..2802)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2791..2807
                     /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)."
     promoter        complement(join(2815..2821,2822..2839,2840..2845))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2860..2881
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2940..3135
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2986..3121
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3141..3275
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3178..3197)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3232..3251
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3343..3360)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3514..4102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3594..3613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4273..5064,5065..5133))
                     /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     4896..4915
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5134..5150,1..88))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
       61 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcga
      121 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      181 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      241 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      301 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      361 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      421 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      481 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
      541 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
      601 ggggcggggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc
      661 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
      721 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
      781 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
      841 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
      901 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc
      961 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg
     1021 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc
     1081 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc
     1141 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
     1201 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc
     1261 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     1321 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1381 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1441 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1501 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1561 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1621 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1681 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1741 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1801 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcat gtcggcgggc
     1861 agcagctgca gccagactcc cagccgggcc atccccactc gccgcgtagc cctcggcgat
     1921 ggcgtgcagc tcccgcccgg ggactacagc accactccgg gcggcacgct cttcagcacc
     1981 accccgggag gaaccaggat tatctatgac cggaaatttc tgatggagtg tcggaactca
     2041 cctgtggcca aaacaccccc aaaggacctg ccagccattc ctggggtcac tagccctacc
     2101 agcgatgagc ctcccatgca agccagccag agccaactgc ccagcagccc ggaagataag
     2161 cgggcaggcg gtgaagagtc acaagctgag atggacattt aagaattcac tcctcaggtg
     2221 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     2281 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     2341 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     2401 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     2461 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     2521 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     2581 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     2641 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     2701 gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta
     2761 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2821 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2881 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg
     2941 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3001 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3061 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3121 ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca
     3181 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3241 ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg
     3301 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3361 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3421 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3481 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3541 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3601 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3661 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3721 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3781 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3841 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3901 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3961 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4021 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4081 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4141 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4201 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4261 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4321 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4381 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4441 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4501 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4561 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4621 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4681 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4741 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4801 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4861 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4921 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4981 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5041 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5101 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
//
