LOCUS       V001188                 8083 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V001188
VERSION     V001188
KEYWORDS    pLJM1-EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8083)
  AUTHORS   Sancak Y, Peterson TR, Shaul YD, Lindquist RA, Thoreen CC, Bar-Peled
            L, Sabatini DM
  TITLE     The Rag GTPases bind raptor and mediate amino acid signaling to
            mTORC1.
  JOURNAL   Science. 2008 Jun 13. 320(5882):1496-501.
   PUBMED   18497260
REFERENCE   2  (bases 1 to 8083)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8083)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science.
            2008 Jun 13. 320(5882):1496-501."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        290..493
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             538..1254
                     /label="EGFP"
                     /note="enhanced GFP"
     misc_feature    1325..1442
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        1491..2001
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             2023..2619
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             2750..2983
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3055..3189
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      3216..3351
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(3372..3390)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3400..3416)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      3558..4013
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4039..4143
                     /label="AmpR promoter"
     CDS             4144..5001
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5175..5763
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     5917..5934
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     protein_bind    6051..6072
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6087..6117
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    6125..6141
                     /label="lac operator"
                     /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     6149..6165
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        6186..6204
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6232..6458
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6459..6639
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6686..6811
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7304..7537
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             7722..7766
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             7915..7956
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
ORIGIN
        1 tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac
       61 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
      121 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgccaagta cgccccctat
      181 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      241 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      301 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      361 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      421 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      481 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccggt cgccaccatg
      541 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      601 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      661 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      721 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
      781 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      841 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      901 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      961 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     1021 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     1081 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1141 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1201 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtccgga
     1261 ctcagatctc gagctcaagc ttcgaattct cgacctcgag acaaatggca gtattcatcc
     1321 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     1381 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     1441 ttcgggttta ttacagggac agcagagatc cactttggcc gcggctcgag ggggttgggg
     1501 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt
     1561 tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc
     1621 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc
     1681 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca
     1741 cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg
     1801 cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag cggccgggaa
     1861 ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg
     1921 tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga
     1981 atcaccgacc tctctcccca gggggatcca ccggagctta ccatgaccga gtacaagccc
     2041 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     2101 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     2161 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     2221 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     2281 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     2341 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     2401 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     2461 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     2521 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     2581 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg
     2641 cagcgcccga ccgaaaggag cgcacgaccc catgcatcgg tacctttaag accaatgact
     2701 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     2761 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     2821 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     2881 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     2941 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     3001 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     3061 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3121 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3181 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3241 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3301 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     3361 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     3421 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     3481 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     3541 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3601 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3661 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3721 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3781 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3841 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3901 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3961 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     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 atgaccatga ttacgccaag
     6181 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt
     6241 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag
     6301 gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat
     6361 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca
     6421 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga
     6481 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     6541 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     6601 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     6661 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     6721 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     6781 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     6841 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     6901 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     6961 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     7021 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     7081 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     7141 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     7201 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     7261 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     7321 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     7381 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     7441 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     7501 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     7561 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     7621 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     7681 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     7741 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     7801 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     7861 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     7921 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     7981 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgatca cgagactagc
     8041 ctcgagcggc cgcccccttc accgagggcc tatttcccat gat
//
