LOCUS       sequence-155339-        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-WT
FEATURES             Location/Qualifiers
     source          1..6267
                     /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"
     CDS             1175..1201
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      2803..2812
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2816..2950
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2853..2872)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2907..2926
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3019..3376
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3019..3210
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3042..3062)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3227..3362
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3396..3413)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3396..3412)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3405..3427)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3625..4080
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3712..3731)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3922..3943
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4097..4116)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4216..4238
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4276..4294)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4362..4466
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4467..4535,4536..5327)
                     /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(4685..4704)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5498..6086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5987..6006
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6240..6257
                     /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 cggttctaag cttctgcagg tcgactctag aatgtaccca tacgatgttc cagattacgc
     1201 tgcaggagtg tttgacatag acctggacca gccagaggat gcaggctctg aggatgagct
     1261 ggaggagggg ggtcagttaa atgaaagcat ggaccatggg ggagttggac catatgaact
     1321 tggcatggaa cattgtgaga aatttgaaat ctcagaaact agtgtgaaca gagggccaga
     1381 aaaaatcaga ccagaatgtt ttgagctact tcgggtactt ggtaaagggg gctatggaaa
     1441 ggtttttcaa gtacgaaaag taacaggagc aaatactggg aagatatttg ccatgaaggt
     1501 gcttaaaaag gcaatgatag taagaaatgc taaagataca gctcatacaa aagcagagcg
     1561 gaatattctg gaggaagtaa agcatccctt catcgtggat ttaatttatg cctttcagac
     1621 cggtggaaaa ctctacctca tccttgagta tctcagtgga ggagaactat ttatgcagtt
     1681 agaaagagag gggatattca tggaagatac agcttgcttt tacttggctg aaatctccat
     1741 ggctttgggg catttacatc aaaaagggat catctacaga gacctgaagc cggagaacat
     1801 catgcttaat caccaaggtc acgtgaagct gacagacttt ggactatgca aagaatctat
     1861 tcatgatgga acagtcacgc acacattttg tggaacaata gaatacatgg cccctgaaat
     1921 cttgatgaga agcggccaca accgtgctgt ggattggtgg agtttgggag cattaatgta
     1981 tgacatgctg actggagcac ctccattcac tggggagaat agaaagaaaa caattgacaa
     2041 aatcctcaaa tgtaaactta atttgcctcc ctacctcaca caagaagctc gagatctgct
     2101 taaaaagctg ctgaaaagaa atgctgcttc tcgtcttgga gctggccctg gggatgctgg
     2161 agaagtccaa gcgcatccat tttttagaca cattaactgg gaagagcttt tggctcggaa
     2221 ggtggagccc ccctttaagc ctctgttgca atctgaagag gatgtgagtc agtttgattc
     2281 aaagtttact cgtcagacac ctgttgacag ccccgatgac tcaactctca gtgaaagtgc
     2341 caaccaggtc tttctgggtt ttacatatgt ggctccatct gtacttgaaa gtgtgaaaga
     2401 aaagttttct tttgaaccaa aaatccgatc gcctcgaaga tttattggta gcccacgaac
     2461 gcctgtcagc ccagtcaaat tctctcctgg ggatttctgg ggacgaggtg cttcagccag
     2521 cacagcaaat cctcagacac ctgtggaata cccaatggaa acaagtggaa tagagcagat
     2581 ggatgtgaca acgagcgggg aagcttcagc gccacttcca atccgacagc ccaactctgg
     2641 gccatacaaa aaacaagctt ttcctatgat ctccaaacgg ccagagcacc tgcgtatgaa
     2701 tctatgatga aacaatgctt ttattaatgc aaatgcaaaa aggaaacaaa tcctggaagg
     2761 ggatgtgtga ggatcccccg ggctgcagga attcaatcga tggccgccat ggcccaactt
     2821 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     2881 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     2941 tgtctggatc gggaattaat tcggcgcagc accatggcct gaaataacct ctgaaagagg
     3001 aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3061 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3121 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3181 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3241 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3301 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3361 cctaggcttt tgcaaaaagc tgttaacagc ttggcactgg ccgtcgtttt acaacgtcgt
     3421 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     3481 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     3541 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3601 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3661 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3721 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3781 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3841 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3901 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3961 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4021 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     4081 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     4141 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     4201 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     4261 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     4321 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4381 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4441 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4501 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4561 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4621 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4681 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4741 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4801 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4861 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4921 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4981 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     5041 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5101 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5161 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5221 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5281 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5341 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5401 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5461 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5521 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5581 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5641 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5701 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5761 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5821 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5881 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5941 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6001 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6061 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6121 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6181 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     6241 gcgagtcagt gagcgaggaa gcggaag
//
