LOCUS       sequence-137768-        5692 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 5692)
  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 5692)
  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 Sec13 pRK5
FEATURES             Location/Qualifiers
     source          1..5692
                     /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      2228..2237
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2241..2375
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2278..2297)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2332..2351
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2444..2801
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2444..2635
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2467..2487)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2652..2787
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2821..2837)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2830..2852)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3050..3505
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3137..3156)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3347..3368
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3522..3541)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3641..3663
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3701..3719)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3787..3891
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3892..3960,3961..4752)
                     /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(4110..4129)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4923..5511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5412..5431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5665..5682
                     /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 attcacgacg cccagatgga ctactatggc acccgcctgg caacctgctc
     1321 atcagacagg tccgtcaaaa tctttgatgt gcgcaatgga gggcagatcc ttatcgccga
     1381 cctcaggggt catgagggtc ctgtgtggca agtggcctgg gctcacccca tgtacggcaa
     1441 catcctggca tcgtgctcct atgaccggaa agtcattatc tggagagagg aaaacggcac
     1501 ctgggagaag agccacgagc atgcgggaca cgactcctca gtgaactcgg tgtgctgggc
     1561 cccccatgac tacggcctga tcctggcctg tgggagctcg gatggggcca tctccctgct
     1621 gacttacacc ggggaaggcc aatgggaagt aaagaagatc aacaacgctc acaccattgg
     1681 ctgcaatgcc gtcagctggg cccctgctgt tgtacctgga agcctcatag accacccatc
     1741 ggggcagaaa cccaattaca tcaagaggtt tgcatcaggt ggctgtgaca acctcatcaa
     1801 gctgtggaag gaggaggagg acggccagtg gaaggaggag cagaagctag aagcgcacag
     1861 tgactgggtt cgagatgtgg cctgggcccc ctccatcggc ctgcccacca gcaccatcgc
     1921 cagctgctcc caggatggtc gtgtgttcat ttggacctgt gatgatgcct caagcaatac
     1981 gtggtcccct aaattgttgc acaagttcaa cgatgtggtg tggcatgtga gctggtccat
     2041 cacagccaac atcctggctg tctctggtgg agacaataag gtgaccctgt ggaaggagtc
     2101 agttgatggg cagtgggtgt gcatcagtga tgtcaacaag ggccagggct ccgtatcagc
     2161 atcagtgaca gagggccagc agaacgagca gtgagcggcc gctaagtaag taaggatccc
     2221 cagcttggcc gccatggccc aacttgttta ttgcagctta taatggttac aaataaagca
     2281 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2341 ccaaactcat caatgtatct tatcatgtct ggatcgggaa ttaattcggc gcagcaccat
     2401 ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc
     2461 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2521 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2581 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2641 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2701 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     2761 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctgtta acagcttggc
     2821 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     2881 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     2941 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     3001 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     3061 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3121 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3181 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3241 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     3301 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3361 caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg
     3421 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3481 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     3541 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3601 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3661 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3721 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3781 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3841 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3901 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3961 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4021 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4081 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     4141 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4201 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4261 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4321 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4381 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4441 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4501 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4561 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4621 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4681 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4741 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4801 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4861 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4921 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4981 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5041 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5101 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5161 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5221 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5281 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5341 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5401 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5461 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5521 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5581 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5641 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga ag
//
