LOCUS       sequence-220590-        5740 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 5740)
  AUTHORS   Kim DH, Sarbassov DD, Ali SM, Latek RR, Guntur KV, Erdjument-Bromage
            H, Tempst P, Sabatini DM
  TITLE     GbetaL, a positive regulator of the rapamycin-sensitive pathway 
            required for the nutrient-sensitive interaction between raptor and 
            mTOR.
  JOURNAL   Mol Cell 2003 Apr;11(4):895-904.
  PUBMED    12718876
REFERENCE   2  (bases 1 to 5740)
  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"; journalName: "Mol Cell"; 
            date: "2003-04"; volume: "11(4)"; pages: "895-904"
COMMENT     Sequence Label: myc GbetaL mutant 6
FEATURES             Location/Qualifiers
     source          1..5740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     539..559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     585..609
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        819..837
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     819..836
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     850..869
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      954..963
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             963..992
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      2031..2040
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2044..2178
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2081..2100)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2135..2154
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2247..2604
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2247..2438
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2270..2290)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2455..2590
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2624..2641)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2624..2640)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2633..2655)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2853..3308
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2940..2959)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3150..3171
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3325..3344)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3444..3466
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3504..3522)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3590..3694
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3695..3763,3764..4555)
                     /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(3913..3932)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4726..5314
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5215..5234
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5468..5485
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5602..5623
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5638..5643,5644..5661,5662..5668)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5676..5692
                     /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     5681..5703
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5700..5716
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5700..5716
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gcccgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
      601 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
      661 cgatccagcc tccgcggccg ggaacggtgc attggaacgc ggattccccg tgccaagagt
      721 gacgtaagta ccgcctatag agtctatagg cccaccccct tggcttcgtt agaacgcggc
      781 tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca ctatagaata
      841 acatccactt tgcctttctc tccacaggtg tccactccca ggtccaactg cacctcggtt
      901 ctatcgattg aattcccgaa ccgacagtcg gtctcttcac caaggccatt cgcgccacca
      961 tggagcaaaa gctcatttct gaggaagatc tcaatggtgg tggtggtggg tcgacgatga
     1021 acacctcccc aggcacggtg ggcagtgacc cggtcatcct ggccactgca ggctacgacc
     1081 acaccgtgcg cttctggcag gcccacagcg gcatctgcac ccggacggtg cagcaccagg
     1141 actcccaggt gaatgccttg gaggtcacac cggaccgcag catgattgct gctgcaggtt
     1201 accagcacat ccgcatgtat gatctcaact ccaataaccc taaccccatc atcagctacg
     1261 acggcgtcaa caagaacatc gcgtctgtgg gcttccacga agacggccgc tggatgtaca
     1321 cgggcggcga ggactgcaca gccaggatct gggacctcag gtcccggaac ctgcagtgcc
     1381 agcggatctt ccaggtgaac gcacccatta actgcgtgtg cctgcacccc aaccaggcag
     1441 agctcatcgt gggtgaccag agcggggcta tccacatctg ggacttgaaa acagaccaca
     1501 acgagcagct gatccctgag cccgaggtct ccatcacgtc cgcccacatc gatcccgacg
     1561 ccagctacat ggcagctgtc aatagcaccg gaaactgcta tgtctggaat ctgacggggg
     1621 gcattggtga cgaggttgat cagctcatcc ccaagactaa gatccctgcc cacacgcgct
     1681 acgccctgca gtgtcgcttc agccccgact ccacgctcct cgccacctgc tcggctgatc
     1741 agacgtgcaa gatctggagg acgtccaact tctccctgat gacggagctg agcatcaaga
     1801 gcggcaaccc cggggagtcc tcccgcggct ggatgtgggg ctgcgccttc tcgggggact
     1861 cccagtacat cgtcactgct tcctcggaca acctggcccg gctctggtgt gtggagactg
     1921 gagagatcaa gagagagtat ggcggccacc agaaggctgt tgtctgcctg gccttcaatg
     1981 acagtgtgct gggctaggcg gccgctaagt aagtaaggat ccccagcttg gccgccatgg
     2041 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2101 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2161 tcttatcatg tctggatcgg gaattaattc ggcgcagcac catggcctga aataacctct
     2221 gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt
     2281 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2341 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2401 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2461 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     2521 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     2581 tttggaggcc taggcttttg caaaaagctg ttaacagctt ggcactggcc gtcgttttac
     2641 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     2701 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2761 gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2821 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     2881 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2941 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3001 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3061 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3121 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3181 caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3241 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     3301 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     3361 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3421 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3481 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3541 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3601 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3661 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3721 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3781 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3841 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3901 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3961 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4021 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4081 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4141 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4201 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4261 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4321 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4381 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4441 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4501 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4561 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4621 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4681 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4741 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4801 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4861 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4921 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4981 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5041 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5101 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5161 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5221 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5281 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5341 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5401 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5461 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     5521 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     5581 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     5641 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     5701 aggaaacagc tatgacatga ttacgaatta attcgagctc
//
