LOCUS       sequence-205959-        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-KR
FEATURES             Location/Qualifiers
     source          1..6267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      19..607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     508..527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     761..778
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    895..916
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(931..936,937..954,955..961)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    969..985
                     /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     974..996
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     993..1009
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     993..1009
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        1038..1417
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1418..1621
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1572..1592
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1618..1642
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1852..1870
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1852..1869
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1883..1902
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1963..1989
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3591..3600
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3604..3738
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3641..3660)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3695..3714
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3807..4164
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3807..3998
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3830..3850)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4015..4150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4184..4201)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4184..4200)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4193..4215)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4413..4868
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4500..4519)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4710..4731
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4885..4904)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5004..5026
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5064..5082)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5150..5254
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5255..5323,5324..6115)
                     /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(5473..5492)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
       61 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      121 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      721 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      781 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
      841 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
      901 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      961 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa
     1021 ttaattcgag ctcgcccgac attgattatt gactagttat taatagtaat caattacggg
     1081 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     1141 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     1201 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     1261 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     1321 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     1381 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     1441 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     1501 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     1561 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     1621 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
     1681 aagacaccgg gaccgatcca gcctccgcgg ccgggaacgg tgcattggaa cgcggattcc
     1741 ccgtgccaag agtgacgtaa gtaccgccta tagagtctat aggcccaccc ccttggcttc
     1801 gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg
     1861 acactataga ataacatcca ctttgccttt ctctccacag gtgtccactc ccaggtccaa
     1921 ctgcacctcg gttctaagct tctgcaggtc gactctagaa tgtacccata cgatgttcca
     1981 gattacgctg caggagtgtt tgacatagac ctggaccagc cagaggatgc aggctctgag
     2041 gatgagctgg aggagggggg tcagttaaat gaaagcatgg accatggggg agttggacca
     2101 tatgaacttg gcatggaaca ttgtgagaaa tttgaaatct cagaaactag tgtgaacaga
     2161 gggccagaaa aaatcagacc agaatgtttt gagctacttc gggtacttgg taaagggggc
     2221 tatggaaagg tttttcaagt acgaaaagta acaggagcaa atactgggaa gatatttgcc
     2281 atgagggtgc ttaaaaaggc aatgatagta agaaatgcta aagatacagc tcatacaaaa
     2341 gcagagcgga atattctgga ggaagtaaag catcccttca tcgtggattt aatttatgcc
     2401 tttcagaccg gtggaaaact ctacctcatc cttgagtatc tcagtggagg agaactattt
     2461 atgcagttag aaagagaggg gatattcatg gaagatacag cttgctttta cttggctgaa
     2521 atctccatgg ctttggggca tttacatcaa aaagggatca tctacagaga cctgaagccg
     2581 gagaacatca tgcttaatca ccaaggtcac gtgaagctga cagactttgg actatgcaaa
     2641 gaatctattc atgatggaac agtcacgcac acattttgtg gaacaataga atacatggcc
     2701 cctgaaatct tgatgagaag cggccacaac cgtgctgtgg attggtggag tttgggagca
     2761 ttaatgtatg acatgctgac tggagcacct ccattcactg gggagaatag aaagaaaaca
     2821 attgacaaaa tcctcaaatg taaacttaat ttgcctccct acctcacaca agaagctcga
     2881 gatctgctta aaaagctgct gaaaagaaat gctgcttctc gtcttggagc tggccctggg
     2941 gatgctggag aagtccaagc gcatccattt tttagacaca ttaactggga agagcttttg
     3001 gctcggaagg tggagccccc ctttaagcct ctgttgcaat ctgaagagga tgtgagtcag
     3061 tttgattcaa agtttactcg tcagacacct gttgacagcc ccgatgactc aactctcagt
     3121 gaaagtgcca accaggtctt tctgggtttt acatatgtgg ctccatctgt acttgaaagt
     3181 gtgaaagaaa agttttcttt tgaaccaaaa atccgatcgc ctcgaagatt tattggtagc
     3241 ccacgaacgc ctgtcagccc agtcaaattc tctcctgggg atttctgggg acgaggtgct
     3301 tcagccagca cagcaaatcc tcagacacct gtggaatacc caatggaaac aagtggaata
     3361 gagcagatgg atgtgacaac gagcggggaa gcttcagcgc cacttccaat ccgacagccc
     3421 aactctgggc catacaaaaa acaagctttt cctatgatct ccaaacggcc agagcacctg
     3481 cgtatgaatc tatgatgaaa caatgctttt attaatgcaa atgcaaaaag gaaacaaatc
     3541 ctggaagggg atgtgtgagg atcccccggg ctgcaggaat tcaatcgatg gccgccatgg
     3601 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3661 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3721 tcttatcatg tctggatcgg gaattaattc ggcgcagcac catggcctga aataacctct
     3781 gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt
     3841 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3901 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3961 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4021 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4081 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4141 tttggaggcc taggcttttg caaaaagctg ttaacagctt ggcactggcc gtcgttttac
     4201 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     4261 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     4321 gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4381 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     4441 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4501 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4561 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4621 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4681 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4741 caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4801 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     4861 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     4921 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4981 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     5041 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     5101 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5161 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5221 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5281 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5341 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5401 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5461 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5521 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5581 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5641 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5701 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5761 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5821 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5881 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5941 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6001 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6061 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6121 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6181 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6241 gttccactga gcgtcagacc ccgtaga
//
