LOCUS       sequence-220600-        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 1
FEATURES             Location/Qualifiers
     source          1..5740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      9..597
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     498..517
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     751..768
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    885..906
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(921..926,927..944,945..951)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    959..975
                     /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     964..986
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     983..999
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     983..999
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        1028..1407
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1408..1611
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1562..1582
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1608..1632
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1842..1860
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1842..1859
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1873..1892
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1977..1986
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1986..2015
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3054..3063
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3067..3201
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3104..3123)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3158..3177
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3270..3627
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3270..3461
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3293..3313)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3478..3613
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3647..3664)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3647..3663)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3656..3678)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3876..4331
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3963..3982)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4173..4194
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4348..4367)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4467..4489
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4527..4545)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4613..4717
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4718..4786,4787..5578)
                     /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(4936..4955)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
       61 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      121 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      181 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      241 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      301 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      361 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      421 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      481 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      541 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      601 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
      661 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
      721 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
      781 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
      841 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      901 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      961 gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa ttaattcgag
     1021 ctcgcccgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     1081 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     1141 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     1201 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     1261 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     1321 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     1381 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     1441 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     1501 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     1561 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
     1621 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
     1681 gaccgatcca gcctccgcgg ccgggaacgg tgcattggaa cgcggattcc ccgtgccaag
     1741 agtgacgtaa gtaccgccta tagagtctat aggcccaccc ccttggcttc gttagaacgc
     1801 ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg acactataga
     1861 ataacatcca ctttgccttt ctctccacag gtgtccactc ccaggtccaa ctgcacctcg
     1921 gttctatcga ttgaattccc gaaccgacag tcggtctctt caccaaggcc attcgcgcca
     1981 ccatggagca aaagctcatt tctgaggaag atctcaatgg tggtggtggt gggtcgacga
     2041 tgaacacctc cccaggcacg gtgggcagtg acccggtcat cctggccact gcaggctacg
     2101 accacaccgt gcgcttctgg caggcccaca gcggcatctg cacccggacg gtgcagcacc
     2161 aggcctccca agtgaatgcc ttggaggtca caccggaccg cagcatgatt gctgctgcag
     2221 gttaccagca catccgcatg tatgatctca actccaataa ccctaacccc atcatcagct
     2281 acgacggcgt caacaagaac atcgcgtctg tgggcttcca cgaagacggc cgctggatgt
     2341 acacgggcgg cgaggactgc acagccagga tctgggacct caggtcccgg aacctgcagt
     2401 gccagcggat cttccaggtg aacgcaccca ttaactgcgt gtgcctgcac cccaaccagg
     2461 cagagctcat cgtgggtgac cagagcgggg ctatccacat ctgggacttg aaaacagacc
     2521 acaacgagca gctgatccct gagcccgagg tctccatcac gtccgcccac atcgatcccg
     2581 acgccagcta catggcagct gtcaatagca ccggaaactg ctatgtctgg aatctgacgg
     2641 ggggcattgg tgacgaggtg acccagctca tccccaagac taagatccct gcccacacgc
     2701 gctacgccct gcagtgtcgc ttcagccccg actccacgct cctcgccacc tgctcggctg
     2761 atcagacgtg caagatctgg aggacgtcca acttctccct gatgacggag ctgagcatca
     2821 agagcggcaa ccccggggag tcctcccgcg gctggatgtg gggctgcgcc ttctcggggg
     2881 actcccagta catcgtcact gcttcctcgg acaacctggc ccggctctgg tgtgtggaga
     2941 ctggagagat caagagagag tatggcggcc accagaaggc tgttgtctgc ctggccttca
     3001 atgacagtgt gctgggctag gcggccgcta agtaagtaag gatccccagc ttggccgcca
     3061 tggcccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3121 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3181 gtatcttatc atgtctggat cgggaattaa ttcggcgcag caccatggcc tgaaataacc
     3241 tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg
     3301 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3361 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3421 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3481 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3541 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3601 ttttttggag gcctaggctt ttgcaaaaag ctgttaacag cttggcactg gccgtcgttt
     3661 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     3721 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     3781 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     3841 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     3901 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3961 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4021 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4081 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4141 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4201 actcaaccct atctcgggct attcttttga tttataaggg attttgccga tttcggccta
     4261 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4321 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4381 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4441 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4501 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4561 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4621 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4681 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4741 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4801 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4861 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4921 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4981 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5041 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5101 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5161 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5221 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5281 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5341 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5401 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5461 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5521 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5581 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5641 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5701 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
//
