LOCUS       sequence-388233-        7699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of kinase-defective MYC-tagged PSK2 (K57A) / TAOK1 (K57A)
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7699)
  AUTHORS   Zihni C, Mitsopoulos C, Tavares IA, Ridley AJ, Morris JD
  TITLE     Prostate-derived sterile 20-like kinase 2 (PSK2) regulates apoptotic
            morphology via C-Jun N-terminal kinase and Rho kinase-1.
  JOURNAL   J Biol Chem. 2006 Mar 17;281(11):7317-23. doi: 
            10.1074/jbc.M513769200. Epub 2006 Jan 2.
  PUBMED    16407310
REFERENCE   2  (bases 1 to 7699)
  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: 
            "10.1074/jbc.M513769200"; journalName: "J Biol Chem"; date: 
            "2006-03-17- 17"; volume: "281"; issue: "11"; pages: "7317-23"
COMMENT     Sequence Label: pRK5-Myc-PSK2 (K57A)
FEATURES             Location/Qualifiers
     source          1..7699
                     /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"
     regulatory      925..934
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             934..963
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3999..4008
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4012..4146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4049..4068)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4103..4122
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4215..4572
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4215..4406
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4238..4258)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4423..4558
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4592..4609)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4592..4608)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4601..4623)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4821..5276
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4908..4927)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5118..5139
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5293..5312)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5412..5434
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5472..5490)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5558..5662
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5663..5731,5732..6523)
                     /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(5881..5900)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6694..7282
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7183..7202
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7436..7453
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7570..7591
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7606..7611,7612..7629,7630..7636)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7644..7660
                     /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     7649..7671
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7668..7684
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7668..7684
                     /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 tatcgattga attggccacc atggagcaga agctgatctc cgaggaggac
      961 ctgggatcca tgccatcaac taacagagca ggcagcctga aggaccctga aattgcagag
     1021 ctcttcttca aagaagatcc agagaagctc ttcacagatc tcagagaaat tggccatgga
     1081 agctttggag cagtgtattt tgcacgagat gtgcgtacca atgaagtggt ggccatcgcg
     1141 aaaatgtctt atagtggaaa gcagtctact gagaaatggc aggatattat taaggaagtc
     1201 aagtttctac aaagaataaa acatcccaac agtatagaat acaaaggctg ttatttacgt
     1261 gaacacacag catggcttgt aatggaatat tgtttaggat ctgcttcgga tttactagaa
     1321 gttcacaaaa agccattaca agaagtggaa atagcagcaa ttacacatgg tgctcttcag
     1381 ggattagcct acttacattc tcatactatg attcatagag atatcaaagc aggaaatatc
     1441 cttctgacag aaccaggcca ggtgaaactt gctgactttg gctctgcttc catggcatca
     1501 cctgccaatt cctttgtggg aacgccgtat tggatggccc cagaagtaat tttagccatg
     1561 gatgaaggac aatatgatgg caaagtagat gtgtggtctc ttggaataac atgtattgaa
     1621 ctagcggaaa ggaagcctcc tttatttaat atgaatgcaa tgagtgcctt atatcacata
     1681 gcccaaaatg aatcccctac actacagtct aatgaatggt ctgattattt tcgcaacttt
     1741 gtagattctt gcctccagaa aatccctcaa gatcgaccta catcagagga acttttaaag
     1801 cacatatttg ttcttcggga gcgccctgaa accgtgttaa tagatctcat tcagaggaca
     1861 aaggatgcag taagagagct ggacaatctg cagtatcgaa agatgaagaa actccttttc
     1921 caggaggcac ataatggacc agcagtagaa gcacaggaag aagaagagga acaagatcat
     1981 ggtgttggcc ggacaggaac agttaatagt gttggaagta atcaatccat tcccagcatg
     2041 tccatcagtg ccagcagcca aagcagtagt gttaacagtc ttccagatgt ctcagatgac
     2101 aagagtgagc tagacatgat ggagggagac cacacagtga tgtctaacag ttctgttatc
     2161 catttaaaac cagaggaaga aaattacaga gaagagggag atcctagaac aagagcatca
     2221 gatccacaat ctccacccca agtatctcgt cacaaatcac actatcgtaa tcgagaacac
     2281 tttgctacta tacggacagc atcactggtt acgaggcaaa tgcaagaaca tgagcaggac
     2341 tctgagctta gagaacaaat gtctggctat aagcgaatga ggcgacaaca tcaaaagcaa
     2401 ctgatgactc tggaaaacaa gctaaaggct gagatggatg aacatcgcct cagattagac
     2461 aaagatcttg aaactcagcg taacaatttt gctgcagaaa tggagaaact tatcaagaaa
     2521 caccaggctg ctatggagaa agaggctaaa gtgatgtcca atgaagagaa aaaatttcag
     2581 caacatattc aggcccaaca gaagaaagaa ctgaatagtt ttctcgagtc ccagaaaaga
     2641 gagtataaac ttcgaaaaga gcagcttaaa gaggagctaa atgaaaacca gagtaccccc
     2701 aaaaaagaaa aacaggagtg gctttcaaag cagaaggaga atatacagca tttccaagca
     2761 gaagaagaag ctaaccttct tcgacgtcaa agacaatacc tagagctgga atgccgtcgc
     2821 ttcaagagaa gaatgttact tgggcgtcat aacttagagc aggaccttgt cagggaggag
     2881 ttaaacaaaa gacagactca gaaggactta gagcatgcca tgctactccg acagcatgaa
     2941 tctatgcaag aactggagtt ccgccacctc aacacaattc agaagatgcg ctgtgagttg
     3001 atcagattac agcatcaaac tgagctcact aaccagctgg aatataataa gcgaagagaa
     3061 cgagaactaa gacgaaagca tgtcatggaa gttcgacaac agcctaagag tttgaagtct
     3121 aaagaactcc aaataaaaaa gcagtttcag gatacctgca aaatccaaac cagacagtac
     3181 aaagcattaa gaaatcacct gctggagact acaccaaaga gtgagcacaa agctgttctg
     3241 aaacggctca aggaggaaca gacccggaaa ttagctatct tggctgagca gtatgatcac
     3301 agcattaatg aaatgctctc cacacaagcc ctgcgtttgg atgaagcaca ggaagcagag
     3361 tgccaggttt tgaagatgca gctgcagcag gaactggagc tgttgaatgc gtatcagagc
     3421 aaaatcaaga tgcaagctga ggcacaacat gatcgagagc ttcgcgagct tgaacagagg
     3481 gtctccctcc ggagggcact cttagaacaa aagattgaag aagagatgtt ggctttgcag
     3541 aatgagcgca cagaacgaat acgaagcctg ttggaacgtc aagccagaga gattgaagct
     3601 tttgactctg aaagcatgag actaggtttt agtaatatgg tcctttctaa tctctcccct
     3661 gaggcattca gccacagcta cccgggagct tctggttggt cacacaaccc tactgggggt
     3721 ccaggacctc actggggtca tcccatgggt ggcccaccac aagcttgggg ccatccaatg
     3781 caaggtggac cccagccatg gggtcaccct tcagggccaa tgcaaggggt acctcgaggt
     3841 agcagtatgg gagtccgcaa tagcccccag gctctgaggc ggacagcttc tgggggacgg
     3901 acagagcagg gcatgagcag aagcacgagt gtcacttcac aaatatccaa tgggtcacac
     3961 atgtcttata cataatctag aattcctgca gaagcttggc cgccatggcc caacttgttt
     4021 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4081 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4141 tggatcggga attaattcgg cgcagcacca tggcctgaaa taacctctga aagaggaact
     4201 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     4261 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4321 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4381 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4441 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4501 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4561 ggcttttgca aaaagctgtt aacagcttgg cactggccgt cgttttacaa cgtcgtgact
     4621 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     4681 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     4741 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4801 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4861 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4921 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4981 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     5041 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5101 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5161 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     5221 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     5281 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     5341 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     5401 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     5461 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5521 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5581 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5641 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5701 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5761 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5821 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5881 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5941 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6001 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6061 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6121 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6181 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6241 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6301 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6361 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6421 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6481 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6541 ctcatatata ctttagattg atttaaaaat tcatttttaa tttaaaagga tctaggtgaa
     6601 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6661 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6721 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6781 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6841 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6901 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6961 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7021 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7081 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7141 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7201 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7261 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7321 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7381 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     7441 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     7501 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     7561 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     7621 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     7681 tgacatgatt acgaattaa
//
