LOCUS       sequence-132858-        6475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6475)
  AUTHORS   Bar-Peled L, Chantranupong L, Cherniack AD, Chen WW, Ottina KA, 
            Grabiner BC, Spear ED, Carter SL, Meyerson M, Sabatini DM
  TITLE     A Tumor suppressor complex with GAP activity for the Rag GTPases 
            that signal amino acid sufficiency to mTORC1.
  JOURNAL   Science. 2013 May 31;340(6136):1100-6. doi: 10.1126/science.1232044.
  PUBMED    23723238
REFERENCE   2  (bases 1 to 6475)
  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: "2013-05-31- 31"; volume: "340"; 
            issue: "6136"; pages: "1100-6"
COMMENT     Sequence Label: HA Nprl3 pRK5
FEATURES             Location/Qualifiers
     source          1..6475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        250..629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        630..833
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     784..804
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     830..854
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1064..1082
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1064..1081
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1095..1114
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1199..1208
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1211..1237
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3011..3020
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3024..3158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3061..3080)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3115..3134
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3227..3584
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3227..3418
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3250..3270)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3435..3570
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3604..3621)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3604..3620)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3613..3635)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3833..4288
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3920..3939)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4130..4151
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4305..4324)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4424..4446
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4484..4502)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4570..4674
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4675..4743,4744..5535)
                     /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(4893..4912)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5706..6294
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6195..6214
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6448..6465
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aattaattcg
      241 agctcgcccg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      301 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      361 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      421 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      481 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      541 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      601 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      661 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      721 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      781 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      841 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      901 gggaccgatc cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca
      961 agagtgacgt aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac
     1021 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata
     1081 gaataacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcacct
     1141 cggttctatc gattgaattc ccgaaccgac agtcggtctc ttcaccaagg ccattcgcgc
     1201 caccatgggc tacccctatg atgtgcctga ctacgcagat ctcaatggtg gtggtggtgg
     1261 gtcgacgatg cgggacaaca ccagccccat cagcgtgatt ctggtgagct cggggagcag
     1321 gggcaataag ctgctgttca ggtacccctt ccagagaagc caggagcacc cggcgtccca
     1381 gacaagtaag ccgcgtagca gatacgctgc cagcaacacg ggcgaccatg ctgatgagca
     1441 ggacggcgat tccaggtttt cagatgttat tctggcaaca attttggcaa ccaagtctga
     1501 aatgtgtggc caaaaatttg aactgaagat tgataatgtg cgatttgttg ggcacccaac
     1561 actgctacag catgctctgg ggcagatctc caaaacagat ccttccccga agagggaagc
     1621 acctactatg attcttttta atgtggtgtt tgcactgagg gccaacgcag acccgtcagt
     1681 gataaactgt ctgcataacc tgtcccgtcg tatcgccacc gtgctgcagc acgaggagcg
     1741 ccgctgccag tacctcaccc gggaggccaa gctgatcctg gcgctccagg atgaggtgtc
     1801 cgccatggct gatggaaatg aaggtcctca gtccccattc catcacatcc tgcccaagtg
     1861 caagctggcc agggacctca aggaagctta tgacagcctg tgcacgtcgg gcgtagttcg
     1921 gcttcacatc aacagctggc tggaggtgag cttctgcctg ccccacaaga tccactatgc
     1981 ggcctccagt ctgatccccc cagaggccat cgaacggagc ctgaaagcca tccgccccta
     2041 ccatgccctg ctgctgctca gtgatgagaa gtccttgctg ggtgagcttc ctattgactg
     2101 ctcccctgcc ctagtgcggg tgatcaagac cacatctgct gtgaagaacc tgcagcagct
     2161 agcccaagat gcggacctgg ccttgctgca ggttttccag cttgcagctc atctggtgta
     2221 ctggggcaag gccatcatca tctacccgct gtgtgagaac aacgtctaca tgctgtctcc
     2281 caatgccagc gtatgtctgt actccccgct ggccgagcag ttctcccacc agttcccatc
     2341 tcatgacctg ccgtccgttc ttgccaagtt ctccttgccg gtctccttgt cagaatttag
     2401 gaatcccctg gcccccgctg tgcaggagac ccagctcatc cagatggtgg tgtggatgct
     2461 gcagcgccgg cttctcatcc agctgcacac ctatgtctgc ctgatggcct cacccagcga
     2521 ggaggagccc cgtccgcgag aggacgacgt ccccttcact gcccgggtcg gcggtcgcag
     2581 cctcagcacg cccaacgccc tcagctttgg ctccccaacc agcagcgatg acatgaccct
     2641 caccagcccc agcatggaca actccagcgc agagctactt cccagcgggg actcgccact
     2701 gaaccagagg atgacggaga acctgctggc cagcctgtcg gagcatgaac gcgcagccat
     2761 cctcagtgta cccgcagccc agaaccctga ggacctccgc atgtttgcca ggctccttca
     2821 ctacttccgc ggacgccacc acctggagga gattatgtac aacgagaaca cgcggcgctc
     2881 ccagctgctc atgctgtttg acaagttccg cagcgtgctg gtggtgacca cccacgagga
     2941 ccctgtcatt gccgtcttcc aggctctgct cccctgagcg gccgctaagt aagtaaggat
     3001 ccccagcttg gccgccatgg cccaacttgt ttattgcagc ttataatggt tacaaataaa
     3061 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3121 tgtccaaact catcaatgta tcttatcatg tctggatcgg gaattaattc ggcgcagcac
     3181 catggcctga aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga
     3241 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3301 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3361 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3421 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3481 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3541 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctg ttaacagctt
     3601 ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     3661 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     3721 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
     3781 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     3841 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3901 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3961 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4021 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     4081 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4141 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     4201 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4261 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     4321 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4381 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4441 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     4501 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4561 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4621 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4681 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4741 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4801 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4861 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4921 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4981 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5041 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5101 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5161 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5221 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5281 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5341 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5401 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5461 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5521 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5581 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5641 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5701 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5761 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5821 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5881 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5941 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6001 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6061 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6121 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6181 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6241 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6301 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6361 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6421 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaag
//
