LOCUS       sequence-139747-        7627 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 7627)
  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 7627)
  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: Cterminal HA WDR59 pRK5
FEATURES             Location/Qualifiers
     source          1..7627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      918..927
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      922..931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3867..3893
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3914..3923
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3927..4061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3964..3983)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4018..4037
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4130..4487
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4130..4321
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4153..4173)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4338..4473
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4507..4524)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4507..4523)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4516..4538)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4736..5191
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4823..4842)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5033..5054
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5208..5227)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5327..5349
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5387..5405)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5473..5577
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5578..5646,5647..6438)
                     /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(5796..5815)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6609..7197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7098..7117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7351..7368
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7485..7506
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7521..7526,7527..7544,7545..7551)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7559..7575
                     /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     7564..7586
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7583..7599
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7583..7599
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgattgaatt cgtcgacgcc accatggcgg cgcgatggag cagcgaaaac gtggttgtag
      961 agttccgtga ctcccaggca actgcgatgt ctgtggactg tcttgggcag catgcagtgc
     1021 tttctggccg cagattctta tacatcgtca atctagatgc ccctttcgaa ggtcaccgaa
     1081 agatctctcg ccagagcaaa tgggacattg gagctgtgca gtggaatcct catgacagct
     1141 ttgcacacta ttttgcggct tcgagtaacc aacgagtaga cctttacaag tggaaagacg
     1201 gcagtgggga agttggcaca accttacaag gccacactcg tgtcatcagc gacttggact
     1261 gggcggtgtt tgagcctgac ctcctggtta ccagctctgt ggacacctac atctacattt
     1321 gggatatcaa agacacaagg aaacctactg ttgcactgtc tgctgttgcg ggtgcctccc
     1381 aggtcaaatg gaataaaaaa aatgctaact gccttgccac cagccatgac ggcgatgtgc
     1441 ggatatggga taagaggaaa cccagtacag cagtggaata tctagccgcc cacctctcca
     1501 aaatccatgg cctggactgg cacccagaca gcgagcacat tcttgctacc tccagtcaag
     1561 acaattctgt gaagttctgg gattaccgcc agcctcggaa atacctcaat attcttcctt
     1621 gccaggtgcc tgtctggaag gccagataca cacctttcag caatggattg gtgactgtga
     1681 tggttcccca gctgcggagg gaaaacagcc ttctcctgtg gaatgtcttt gacttgaaca
     1741 ccccagtcca caccttcgtg gggcatgatg atgtggtcct ggagttccag tggaggaagc
     1801 agaaggaagg gtccaaggac tatcaactgg tgacgtggtc ccgggatcag accttgagaa
     1861 tgtggcgggt ggattcccag atgcagaggc tttgtgcaaa tgacatatta gatggtgttg
     1921 atgagttcat tgagagtatt tcccttctgc cggaacctga gaagaccctg cacactgaag
     1981 atacagatca ccagcacact gcaagccatg gggaggaaga agccctaaaa gaagatcccc
     2041 ctagaaatct cctggaagag aggaaatcag atcaactggg gctgcctcag accttgcagc
     2101 aggaattctc cctgatcaat gtgcaaatcc ggaatgtcaa tgtggagatg gatgcggcag
     2161 acaggagctg cacagtgtct gtgcactgca gcaaccatcg tgtcaagatg ctggtgaagt
     2221 tccctgcaca gtacccaaac aacgccgccc cttccttcca gtttattaac cccacaacca
     2281 tcacatccac catgaaagct aagctgctga agatcctgaa ggacacagcc ctgcagaaag
     2341 tgaagcgtgg ccagagctgc ctggagccct gcctgcgcca gctcgtctcc tgccttgagt
     2401 cctttgtgaa ccaggaagac agcgcttcca gcaacccgtt tgcactcccc aactctgtca
     2461 ctcccccctt accgacgttt gcgcgggtga ccacggctta cgggtcgtac caggacgcca
     2521 acattccctt tcctaggact tctggggcca ggttctgcgg agcaggttac ctggtatatt
     2581 tcacaaggcc catgacaatg catcgggcgg tgtctcccac agagcctact ccgagatctc
     2641 tctcagcctt gtctgcttat cacactggct tgatcgcgcc catgaagatc cgcacagagg
     2701 cccctgggaa ccttcgttta tacagtggga gccccactcg cagcgagaaa gagcaggtct
     2761 ccatcagctc cttctactac aaggagcgga aatcaagacg atggaaaagt aagcgtgagg
     2821 gatcagactc tggcaatcga cagatcaagg ctgctgggaa agtcatcatc caggatattg
     2881 cttgcctcct gcctgttcac aaatcgctgg gagagctgta catattgaat gtgaatgata
     2941 ttcaggaaac atgtcagaag aatgccgcct ctgccttgct cgttggaaga aaggatcttg
     3001 tccaggtttg gtcgctggct acggtagcta cagatctttg ccttggtccg aaatctgacc
     3061 cagatttgga aacaccctgg gctcgacatc catttgggcg gcagctgctg gagtccctgt
     3121 tggctcacta ttgccggctc cgggatgttc agacactggc gatgctctgt agcgtgtttg
     3181 aagcccagtc tcggcctcag gggctaccaa acccctttgg gccttttcct aaccgttctt
     3241 ctaatcttgt ggtgtcccat agtcgatatc ctagctttac ctcttctggt tcctgctcca
     3301 gtatgtcaga cccagggctc aacactggcg gctggaacat agcgggaaga gaggcagagc
     3361 acttgtcctc cccttgggga gaatcctcac cagaagagct ccgctttggg agtctgacct
     3421 acagtgatcc ccgtgagcga gaacgcgacc agcatgataa aaataaaagg ctcctggacc
     3481 ccgccaatac ccagcaattt gatgacttta agaaatgcta tggggaaatc ctctaccgtt
     3541 ggggtctgag agagaagcga gctgaagtgt tgaagtttgt ctcctgtcct cctgaccctc
     3601 acaaagggat cgagttcggc gtgtactgca gccactgccg gagtgaggtc cgtggcacgc
     3661 agtgtgccat ctgcaaaggc ttcacgttcc agtgtgccat ctgtcacgtg gctgtgcggg
     3721 gatcgtccaa tttctgcctg acctgtgggc acggtggcca caccagccac atgatggagt
     3781 ggtttcggac ccaggaggtg tgtcccaccg ggtgtgggtg ccactgcctg cttgaaagca
     3841 ctttcgcggc cgcaggaggt ggaggttacc cctatgatgt gcctgactac gcatgataag
     3901 gatccccagc ttggccgcca tggcccaact tgtttattgc agcttataat ggttacaaat
     3961 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4021 gtttgtccaa actcatcaat gtatcttatc atgtctggat cgggaattaa ttcggcgcag
     4081 caccatggcc tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa
     4141 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4201 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4261 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4321 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4381 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4441 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctgttaacag
     4501 cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     4561 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     4621 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt
     4681 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     4741 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4801 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4861 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4921 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     4981 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5041 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg
     5101 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5161 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     5221 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     5281 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     5341 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     5401 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5461 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5521 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5581 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5641 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5701 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5761 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5821 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5881 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5941 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6001 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6061 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6121 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6181 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6241 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6301 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6361 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6421 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6481 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6541 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6601 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6661 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6721 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6781 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6841 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6901 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6961 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7021 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7081 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7141 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7201 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7261 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7321 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7381 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     7441 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7501 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7561 gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa ttaattcgag
     7621 ctcgccc
//
