LOCUS       sequence-142540-        6958 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 6958)
  AUTHORS   Richards SA, Dreisbach VC, Murphy LO, Blenis J
  TITLE     Characterization of regulatory events associated with membrane 
            targeting of p90 ribosomal S6 kinase 1.
  JOURNAL   Mol Cell Biol. 2001 Nov . 21(21):7470-80.
  PUBMED    11585927
REFERENCE   2  (bases 1 to 6958)
  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: "Mol Cell 
            Biol. 2001 Nov . 21(21):7470-80."
COMMENT     Sequence Label: pKH3-human RSK1
FEATURES             Location/Qualifiers
     source          1..6958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             939..965
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             972..998
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1005..1031
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3258..3267
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3271..3405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3308..3327)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3362..3381
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3474..3831
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3474..3665
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3497..3517)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3682..3817
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3851..3868)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3851..3867)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3860..3882)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4080..4535
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4167..4186)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4377..4398
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4552..4571)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4671..4693
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4731..4749)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4817..4921
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4922..4990,4991..5782)
                     /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(5140..5159)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5953..6541
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6442..6461
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6695..6712
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6829..6850
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6865..6870,6871..6888,6889..6895)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6903..6919
                     /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     6908..6930
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6927..6943
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6927..6943
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc taagcttctg caggtcgact ctagaatgta cccatacgat gttccagatt
      961 acgctggatc ttacccatac gatgttccag attacgctgg atcttaccca tacgatgttc
     1021 cagattacgc tggatccgaa ttcatgccgc tcgcccagct caaggagccc tggccgctca
     1081 tggagctagt gccgctggac ccggagaatg gacagacctc aggggaagaa gctggacttc
     1141 agccgtccaa ggatgagggc gtcctcaagg agatctccat cacgcaccac gtcaaggctg
     1201 gctctgagaa ggctgatcca tcccatttcg agctcctcaa ggttctgggc cagggatcct
     1261 ttggcaaagt cttcctggtg cggaaagtca cccggcctga cagtgggcac ctgtatgcta
     1321 tgaaggtgct gaagaaggca acgctgaaag tacgtgaccg cgtccggacc aagatggaga
     1381 gagacatcct ggctgatgta aatcacccat tcgtggtgaa gctgcactat gccttccaga
     1441 ccgagggcaa gctctatctc attctggact tcctgcgtgg tggggacctc ttcacccggc
     1501 tctcaaaaga ggtgatgttc acggaggagg atgtgaagtt ttacctggcc gagctggctc
     1561 tgggcctgga tcacctgcac agcctgggta tcatttacag agacctcaag cctgagaaca
     1621 tccttctgga tgaggagggc cacatcaaac tcactgactt tggcctgagc aaagaggcca
     1681 ttgaccacga gaagaaggcc tattctttct gcgggacagt ggagtacatg gcccctgagg
     1741 tcgtcaaccg ccagggccac tcccatagtg cggactggtg gtcctatggg gtgttgatgt
     1801 ttgagatgct gacgggctcc ctgcccttcc aggggaagga ccggaaggag accatgacac
     1861 tgattctgaa ggcgaagcta ggcatgcccc agtttctgag cactgaagcc cagagcctct
     1921 tgcgggccct gttcaagcgg aatcctgcca accggctcgg ctccggccct gatggggcag
     1981 aggaaatcaa gcggcatgtc ttctactcca ccattgactg gaataagcta taccgtcgtg
     2041 agatcacgcc acccttcaag ccagcagtgg ctcagcctga tgacaccttc tactttgaca
     2101 ccgagttcac gtcccgcaca cccaaggatt ccccaggcat cccccccagc gctggggccc
     2161 atcagctgtt ccggggcttc agcttcgtgg ccaccggcct gatggaagac gacggcaagc
     2221 ctcgtgcccc gcaggcaccc ctgcactcgg tggtacagca actccatggg aagaacctgg
     2281 tttttagtga cggctacgtg gtaaaggaga caattggtgt gggctcctac tctgagtgca
     2341 agcgctgtgt ccacaaggcc accaacatgg agtatgctgt caaggtcatt gataagagca
     2401 agcgggatcc ttcagaagag attgagattc ttctgcggta tggccagcac cccaacatca
     2461 tcactctgaa agatgtgtat gatgatggca aacacgtgta cctggtgaca gagctgatgc
     2521 ggggtgggga gctgctggac aagatcctgc ggcagaagtt cttctcagag cgggaggcca
     2581 gctttgtcct gcacaccatt ggcaaaactg tggagtatct gcactcacag ggggttgtgc
     2641 acagggacct gaagcccagc aacatcctgt atgtggacga gtccgggaat cccgagtgcc
     2701 tgcgcatctg tgactttggt tttgccaaac agctgcgggc tgagaatggg ctcctcatga
     2761 caccttgcta cacagccaac tttgtggcgc ctgaggtgct gaagcgccag ggctacgatg
     2821 aaggctgcga catctggagc ctgggcattc tgctgtacac catgctggca ggatatactc
     2881 catttgccaa cggtcccagt gacacaccag aggaaatcct aacccggatc ggcagtggga
     2941 agtttaccct cagtggggga aattggaaca cagtttcaga gacagccaag gacctggtgt
     3001 ccaagatgct acacgtggat ccccaccagc gcctcacagc taagcaggtt ctgcagcatc
     3061 catgggtcac ccagaaagac aagcttcccc aaagccagct gtcccaccag gacctacagc
     3121 ttgtgaaggg agccatggct gccacgtact ccgcactcaa cagctccaag cccacccccc
     3181 agctgaagcc catcgagtca tccatcctgg cccagcggcg agtgaggaag ttgccatcca
     3241 ccaccctgtg atcgatggcc gccatggccc aacttgttta ttgcagctta taatggttac
     3301 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3361 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcgggaa ttaattcggc
     3421 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc
     3481 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3541 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3601 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3661 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3721 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3781 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctgtta
     3841 acagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     3901 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     3961 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     4021 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     4081 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4141 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4201 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4261 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     4321 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4381 actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata
     4441 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4501 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     4561 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     4621 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4681 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4741 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4801 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4861 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4921 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4981 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5041 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5101 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5161 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5221 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5281 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5341 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5401 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5461 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5521 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5581 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5641 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5701 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5761 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5821 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5881 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5941 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6001 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6061 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6121 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6181 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6241 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6301 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6361 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6421 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6481 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6541 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6601 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     6661 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     6721 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     6781 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     6841 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     6901 aattgtgagc ggataacaat ttcacacagg aaacagctat gacatgatta cgaattaa
//
