LOCUS       sequence-424510-        7138 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 7138)
  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 7138)
  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 WDR24 pRK5
FEATURES             Location/Qualifiers
     source          1..7138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      141..276
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(310..327)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(310..326)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(319..341)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      539..994
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(626..645)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     836..857
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1011..1030)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1130..1152
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1190..1208)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1276..1380
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1381..1449,1450..2241)
                     /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(1599..1618)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2412..3000
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2901..2920
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3154..3171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3288..3309
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3324..3329,3330..3347,3348..3354)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3362..3378
                     /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     3367..3389
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3386..3402
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3386..3402
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        3431..3810
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3811..4014
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3965..3985
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4011..4035
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        4245..4263
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4245..4262
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     4276..4295
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      4380..4389
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4392..4418
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      6855..6864
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    6868..7002
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6905..6924)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6959..6978
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        join(7071..7138,1..290)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        join(7071..7138,1..124)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7094..7114)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
       61 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
      121 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
      181 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
      241 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctgttaa
      301 cagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
      361 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
      421 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
      481 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
      541 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
      601 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
      661 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
      721 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
      781 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
      841 ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa
      901 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
      961 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     1021 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     1081 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     1141 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     1201 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     1261 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     1321 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     1381 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     1441 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     1501 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     1561 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     1621 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     1681 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     1741 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     1801 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     1861 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     1921 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     1981 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     2041 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     2101 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     2161 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     2221 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     2281 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     2341 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     2401 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2461 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2521 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     2581 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2641 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2701 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2761 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2821 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2881 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2941 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3001 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3061 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3181 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     3241 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     3301 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     3361 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaattaattc
     3421 gagctcgccc gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     3481 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     3541 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     3601 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     3661 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     3721 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     3781 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     3841 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     3901 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     3961 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
     4021 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     4081 cgggaccgat ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc
     4141 aagagtgacg taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa
     4201 cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat
     4261 agaataacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc
     4321 tcggttctat cgattgaatt cccgaaccga cagtcggtct cttcaccaag gccattcgcg
     4381 ccaccatggg ctacccctat gatgtgcctg actacgcaga tctcaatggt ggtggtggtg
     4441 ggtcgacgat ggagaagatg tcccgtgtga ccacagccct gggtggcagc gtgctgacag
     4501 gccgcaccat gcactgccac ctggatgctc ccgccaatgc catcagtgtg tgccgcgacg
     4561 cagcccaggt ggtcgtggca ggccgtagca tcttcaagat ctatgccatc gaggaggaac
     4621 agttcgtgga aaagctgaac ctgcgtgtgg ggcgcaagcc ttcgcttaac ctgagctgtg
     4681 ctgacgtggt ctggcaccag atggatgaga acctgctggc cacagcagcc accaatggcg
     4741 tggtggtcac gtggaacctg ggccggccat cccgcaacaa gcaggaccag ctgttcacag
     4801 aacacaagcg cacggtaaac aaagtctgct tccaccccac cgaagcccac gtgctgctca
     4861 gtggctccca ggatggcttc atgaagtgct ttgacctccg cagaaaggac tctgtcagca
     4921 ccttctcggg ccagtcggag agcgtgcggg acgtgcagtt cagtatccgg gactacttca
     4981 ccttcgcctc cacctttgag aacggcaatg tgcagctctg ggacatccgg cgtcccgacc
     5041 ggtgcgagag gatgttcaca gcccacaacg gacccgtctt ctgctgcgac tggcaccccg
     5101 aggacagggg ctggttggcc actggagggc gcgacaagat ggtgaaggtc tgggacatga
     5161 ccacgcaccg tgccaaggag atgcactgtg tgcagaccat cgcctcggtg gcccgtgtga
     5221 agtggcggcc agagtgccgc caccacctgg ccacgtgctc catgatggtg gaccacaaca
     5281 tctatgtttg ggacgtgcgc cggcccttcg tgccagctgc catgtttgag gaacaccgag
     5341 acgtcaccac gggaattgcc tggcgccacc cccacgaccc ctccttcctg ctgtctggct
     5401 ccaaggacag ctcgctgtgc cagcacctgt tccgcgacgc cagccagccc gtcgagcgcg
     5461 ccaaccctga gggcctctgc tacggcctct tcggggacct ggccttcgcc gccaaggaga
     5521 gcctcgtggc tgccgagtcg gggcgcaagc cctacactgg cgaccggcgc caccccatct
     5581 tctttaagcg caagctggac cctgccgagc ccttcgcagg cctcgcctcc agtgccctca
     5641 gtgtctttga gacggagcca ggtggcggcg gcatgcgctg gtttgtggac acagctgagc
     5701 gttatgcgct ggctggccgg ccactggccg agctctgtga ccacaacgca aaggtggctc
     5761 gagagcttgg ccgcaaccag gtggcgcaaa cgtggaccat gctgcggatc atctactgca
     5821 gccctggcct agtgcccact gcaaacctca accacagtgt gggcaagggt ggctcctgtg
     5881 gcctcccgct catgaacagt ttcaacctga aggatatggc cccagggttg ggcagtgaga
     5941 cgcggctgga ccgcagcaaa ggagatgcgc ggagcgacac agttctgctc gactcctcgg
     6001 ccacactcat caccaatgag gataacgagg aaaccgaggg cagcgacgta cctgccgact
     6061 acctgctggg tgacgtggaa ggtgaggagg acgagctgta cctgctggat ccggaacacg
     6121 cgcaccccga ggaccctgag tgcgtgctgc cgcaggaggc ctttccgctg cgccacgaga
     6181 tcgtggacac gcctcccggg cccgagcacc tgcaggacaa ggccgactcc ccgcacgtga
     6241 gcggcagcga ggcggatgtg gcctccctgg cccccgtgga ctcctccttc tcgctcctgt
     6301 ctgtctcaca cgcgctctac gacagccgcc tgccgcccga cttcttcggc gtgctggtgc
     6361 gcgacatgct gcacttctac gctgagcagg gcgacgtgca gatggctgtg tctgtgctca
     6421 tcgtcctggg tgaacgggtg cgcaaggaca tcgacgagca gacccaggag cactggtaca
     6481 cttcctacat cgacctgctg cagcgcttcc gcctctggaa cgtgtccaac gaggtggtca
     6541 agctgagcac cagccgcgcc gtcagctgcc tcaaccaggc ctccaccacc ctgcacgtca
     6601 actgcagcca ctgcaagcgg cccatgagca gccggggctg ggtctgcgac aggtgccacc
     6661 gctgcgccag catgtgtgcc gtctgccacc acgtagtcaa gggtctcttc gtgtggtgcc
     6721 agggctgcag ccacggcggc cacctgcagc acatcatgaa gtggctggaa ggcagctccc
     6781 actgtcccgc aggctgcggc cacctctgcg agtactcctg agcggccgct aagtaagtaa
     6841 ggatccccag cttggccgcc atggcccaac ttgtttattg cagcttataa tggttacaaa
     6901 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6961 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcgggaatta attcggcgca
     7021 gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
     7081 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccag
//
