LOCUS       sequence-409931-        6267 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 6267)
  AUTHORS   Schalm SS, Blenis J
  TITLE     Identification of a conserved motif required for mTOR signaling.
  JOURNAL   Curr Biol. 2002 Apr 16. 12(8):632-9.
  PUBMED    11967149
REFERENCE   2  (bases 1 to 6267)
  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: "Curr Biol. 
            2002 Apr 16. 12(8):632-9."
COMMENT     Sequence Label: pRK7-HA-S6K1-F5A-E389-R3A
FEATURES             Location/Qualifiers
     source          1..6267
                     /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"
     CDS             926..952
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      2554..2563
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2567..2701
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2604..2623)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2658..2677
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2770..3127
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2770..2961
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2793..2813)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2978..3113
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3147..3164)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3147..3163)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3156..3178)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3376..3831
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3463..3482)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3673..3694
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3848..3867)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3967..3989
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4027..4045)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4113..4217
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4218..4286,4287..5078)
                     /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(4436..4455)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5249..5837
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5738..5757
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5991..6008
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6125..6146
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6161..6166,6167..6184,6185..6191)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6199..6215
                     /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     6204..6226
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6223..6239
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6223..6239
                     /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 tcggttctaa
      901 gcttctgcag gtcgactcta gaatgtaccc atacgatgtt ccagattacg ctgcaggagt
      961 ggctgacata gacctggacc agccagagga tgcaggctct gaggatgagc tggaggaggg
     1021 gggtcagtta aatgaaagca tggaccatgg gggagttgga ccatatgaac ttggcatgga
     1081 acattgtgag aaatttgaaa tctcagaaac tagtgtgaac agagggccag aaaaaatcag
     1141 accagaatgt tttgagctac ttcgggtact tggtaaaggg ggctatggaa aggtttttca
     1201 agtacgaaaa gtaacaggag caaatactgg gaagatattt gccatgaagg tgcttaaaaa
     1261 ggcaatgata gtaagaaatg ctaaagatac agctcataca aaagcagagc ggaatattct
     1321 ggaggaagta aagcatccct tcatcgtgga tttaatttat gcctttcaga ccggtggaaa
     1381 actctacctc atccttgagt atctcagtgg aggagaacta tttatgcagt tagaaagaga
     1441 ggggatattc atggaagata cagcttgctt ttacttggct gaaatctcca tggctttggg
     1501 gcatttacat caaaaaggga tcatctacag agacctgaag ccggagaaca tcatgcttaa
     1561 tcaccaaggt cacgtgaagc tgacagactt tggactatgc aaagaatcta ttcatgatgg
     1621 aacagtcacg cacacatttt gtggaacaat agaatacatg gcccctgaaa tcttgatgag
     1681 aagcggccac aaccgtgctg tggattggtg gagtttggga gcattaatgt atgacatgct
     1741 gactggagca cctccattca ctggggagaa tagaaagaaa acaattgaca aaatcctcaa
     1801 atgtaaactt aatttgcctc cctacctcac acaagaagct cgagatctgc ttaaaaagct
     1861 gctgaaaaga aatgctgctt ctcgtcttgg agctggccct ggggatgctg gagaagtcca
     1921 agcgcatcca ttttttagac acattaactg ggaagagctt ttggctcgga aggtggagcc
     1981 cccctttaag cctctgttgc aatctgaaga ggatgtgagt cagtttgatt caaagtttac
     2041 tcgtcagaca cctgttgaca gccccgatga ctcaactctc agtgaaagtg ccaaccaggt
     2101 ctttctgggt ttcgaatatg tggctccatc tgtacttgaa agtgtgaaag aaaagttttc
     2161 ttttgaacca aaaatcgcat cgcctgcagc atttattggt agcccacgaa cgcctgtcag
     2221 cccagtcaaa ttctctcctg gggatttctg gggacgaggt gcttcagcca gcacagcaaa
     2281 tcctcagaca cctgtggaat acccaatgga aacaagtgga atagagcaga tggatgtgac
     2341 aacgagcggg gaagcttcag cgccacttcc aatccgacag cccaactctg ggccatacaa
     2401 aaaacaagct tttcctatga tctccaaacg gccagagcac ctgcgtatga atctatgatg
     2461 aaacaatgct tttattaatg caaatgcaaa aaggaaacaa atcctggaag gggatgtgtg
     2521 aggatccccc gggctgcagg aattcaatcg atggccgcca tggcccaact tgtttattgc
     2581 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     2641 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     2701 cgggaattaa ttcggcgcag caccatggcc tgaaataacc tctgaaagag gaacttggtt
     2761 aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     2821 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2881 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2941 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3001 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3061 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3121 ttgcaaaaag ctgttaacag cttggcactg gccgtcgttt tacaacgtcg tgactgggaa
     3181 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     3241 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     3301 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     3361 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3421 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3481 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3541 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     3601 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3661 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     3721 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3781 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     3841 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     3901 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3961 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     4021 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4081 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4141 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4201 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4261 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4321 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4381 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4441 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4501 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4561 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4621 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4681 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4741 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4801 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4861 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4921 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4981 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5041 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5101 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5161 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5221 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5281 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5341 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5401 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5461 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5521 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5581 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5641 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5701 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5761 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5821 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5881 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5941 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6001 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     6061 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     6121 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     6181 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca
     6241 tgattacgaa ttaattcgag ctcgccc
//
