LOCUS       sequence-388270-        8442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of kinase-deficient MYC-tagged PSK1-alpha (K57A) / TAOK2 
            (K57A) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8442)
  AUTHORS   Moore TM, Garg R, Johnson C, Coptcoat MJ, Ridley AJ, Morris JD
  TITLE     PSK, a novel STE20-like kinase derived from prostatic carcinoma that
            activates the c-Jun N-terminal kinase mitogen-activated protein 
            kinase pathway and regulates actin cytoskeletal organization.
  JOURNAL   J Biol Chem. 2000 Feb 11;275(6):4311-22. doi: 
            10.1074/jbc.275.6.4311.
  PUBMED    10660600
REFERENCE   2  (bases 1 to 8442)
  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.275.6";
            journalName: "J Biol Chem"; date: "2000-02-11- 11"; volume: "275"; 
            issue: "6"; pages: "4311-22"
COMMENT     Sequence Label: pRK5 Myc-tagged PSK (K57A)
FEATURES             Location/Qualifiers
     source          1..8442
                     /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      4742..4751
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4755..4889
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4792..4811)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4846..4865
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4958..5315
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4958..5149
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4981..5001)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5166..5301
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5335..5352)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5335..5351)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5344..5366)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5564..6019
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5651..5670)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5861..5882
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6036..6055)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6155..6177
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6215..6233)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6301..6405
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6406..6474,6475..7266)
                     /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(6624..6643)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7437..8025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7926..7945
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8179..8196
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8313..8334
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8349..8354,8355..8372,8373..8379)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8387..8403
                     /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     8392..8414
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8411..8427
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8411..8427
                     /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 tgccagctgg gggccgggcc gggagcctga aggacccaga tgtggctgag
     1021 ctcttcttca aggatgaccc agaaaagctc ttctctgacc tccgggaaat tggccatggc
     1081 agctttggag ccgtatactt tgcccgggat gtccggaata gtgaggtggt ggccatcgcg
     1141 aagatgtcct acagtgggaa gcagtccaat gagaaatggc aagacatcat caaggaggtg
     1201 cggttcttac agaagctccg gcatcccaac accattcagt accggggctg ttacctgagg
     1261 gagcacacgg cttggctggt aatggagtat tgcctgggct cagcttctga ccttctagaa
     1321 gtgcacaaga aaccccttca ggaggtagag atcgcagctg tgacccacgg ggcgcttcag
     1381 ggcctggcat atctgcactc ccacaacatg atccataggg atgtgaaggc tggaaacatc
     1441 ctgctgtcag agccagggtt agtgaagcta ggggactttg gttctgcgtc catcatggca
     1501 cctgccaact ccttcgtggg caccccatac tggatggcac ccgaggtgat cctggccatg
     1561 gatgaggggc agtacgatgg caaagtggac gtctggtcct tggggataac ctgcatcgag
     1621 ctggctgaac ggaaaccacc gctctttaac atgaatgcga tgagtgcctt ataccacatt
     1681 gcacagaacg aatcccccgt gctccagtca ggacactggt ctgagtactt ccggaatttt
     1741 gtcgactcct gtcttcagaa aatccctcaa gacagaccaa cctcagaggt tctcctgaag
     1801 caccgctttg tgctccggga gcggccaccc acagtcatca tggacctgat ccagaggacc
     1861 aaggatgccg tgcgggagct ggacaacctg cagtaccgca agatgaagaa gatcctgttc
     1921 caagaggcac ccaacggccc tggtgccgag gccccagagg aggaagagga ggccgagccc
     1981 tacatgcacc gggccgggac tctgaccagc ctcgagagta gccactcagt gcccagcatg
     2041 tccatcagcg cctccagcca gagcagctcc gtcaacagcc tagcagatgc ctcagacaac
     2101 gaggaagagg aggaggagga ggaggaagag gaggaggagg aagaaggccc tgaagcccgg
     2161 gagatggcca tgatgcagga gggggagcac acagtcacct ctcacagctc cattatccac
     2221 cggctgccgg gctctgacaa cctatatgat gacccctacc agccagagat aacccccagc
     2281 cctctccagc cgcctgcagc cccagctccc acttccacca cctcttccgc ccgccgccgg
     2341 gcctactgcc gtaaccgaga ccactttgcc accatccgaa ccgcctccct ggtcagccgt
     2401 cagatccagg agcatgagca ggactctgcg ctgcgggagc agctgagcgg ctataagcgg
     2461 atgcgacgac agcaccagaa gcagctgctg gccctggagt cacggctgag gggtgaacgg
     2521 gaggagcaca gtgcacggct gcagcgggag cttgaggcgc agcgggctgg ctttggggca
     2581 gaggcagaaa agctggcccg gcggcaccag gccataggtg agaaggaggc acgagctgcc
     2641 caggccgagg agcggaagtt ccagcagcac atccttgggc agcagaagaa ggagctggct
     2701 gccctgctgg aggcacagaa gcggacctac aaacttcgca aggaacagct gaaggaggag
     2761 ctccaggaga accccagcac tcccaagcgg gagaaggccg agtggctgct gcggcagaag
     2821 gagcagctcc agcagtgcca ggcggaggag gaagcagggc tgctgcggcg gcagcgccag
     2881 tactttgagc tgcagtgtcg ccagtacaag cgcaagatgt tgctggctcg gcacagcctg
     2941 gaccaggacc tgctgcggga ggacctgaac aagaagcaga cccagaagga cttggagtgt
     3001 gcactgctgc ttcggcagca cgaggccacg cgggagctgg agctgcggca gctccaggcc
     3061 gtgcagcgca cgcgggctga gctcacccgc ctgcagcacc agacggagct gggcaaccag
     3121 ctggagtaca acaagcggcg tgagcaagag ttgcggcaga agcatgcggc ccaggttcgc
     3181 cagcagccca agagcctcaa agtacgtgca ggccagcgcc ccccgggcct tccactcccc
     3241 attcctgggg ctctgggccc acccaacaca ggcaccccta tagaacagca gccctgctca
     3301 cctggccagg aggcagtcct ggaccaaaga atgcttggcg aggaggagga agcagttgga
     3361 gagagaagga ttctgggaaa ggaaggggcc actttggagc ccaagcagca gaggattctg
     3421 ggggaagaat caggagcccc tagtcccagt ccacaaaaac atgggagcct ggttgatgag
     3481 gaagtttggg gtctgcctga ggagatagag gagcttaggg tgccctccct tgtaccccag
     3541 gagaggagca ttgttggcca ggaggaggct gggacgtgga gcttgtgggg gaaggaggat
     3601 gagagtcttc tggatgagga gtttgagctt ggctgggtcc agggcccagc actgactccc
     3661 gtccctgagg aggaggaaga agaggaagag ggggctccga ttgggacccc tagggatcct
     3721 ggagatggtt gtccttcccc cgacatccct cctgaacccc ctccaacaca cctgaggccc
     3781 tgccctgcca gccagctccc tggactcctg tcccatggcc tcctggccgg cctctccttt
     3841 gcagtggggt cctcctctgg cctcctgccc ctcctgctgc tgctgctgct tccattgctg
     3901 gcagcccagg gtgggggtgg cctgcaggca gcgctgctgg cccttgaggt ggggctggtg
     3961 ggtctggggg cctcctacct gctcctttgt acagccctgc acctgccctc cagtcttttc
     4021 ctactcctgg cccagggtac cgcactgggg gccgtcctgg gcctgagctg gcgccgaggc
     4081 ctcatgggtg ttcccctggg ccttggagct gcctggctct tagcttggcc aggcctagct
     4141 ctacctctgg tggctatggc agcggggggc agatgggtgc ggcagcaggg cccccgggtg
     4201 cgccggggca tatctcgact ctggttgcgg gttctgctgc gcctgtcacc catggccttc
     4261 cgggccctgc agggctgtgg ggctgtgggg gaccggggtc tgtttgcact gtaccccaaa
     4321 accaacaagg atggcttccg cagccgcctg cccgtccctg ggccccggcg gcgtaatccc
     4381 cgcaccaccc aacacccatt agctctgttg gcaagggtct gggtcctgtg caagggctgg
     4441 aactggcgtc tggcacgggc cagccagggt ttagcatccc acttgccccc gtgggccatc
     4501 cacacactgg ccagctgggg cctgcttcgg ggtgaacggc ccacccgaat cccccggcta
     4561 ctaccacgca gccagcgcca gctagggccc cctgcctccc accagccact gccagggact
     4621 ctagccgggc ggaggtcacg cacccgccag tcccgggccc tgcccccctg gaggtagctg
     4681 actccagccc ttccagccca aatctagagc attgagtaga tagaattcct gcagaagctt
     4741 ggccgccatg gcccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4801 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4861 tcatcaatgt atcttatcat gtctggatcg ggaattaatt cggcgcagca ccatggcctg
     4921 aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag aaccagctgt
     4981 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5041 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     5101 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     5161 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5221 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     5281 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct gttaacagct tggcactggc
     5341 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     5401 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     5461 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
     5521 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     5581 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5641 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5701 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5761 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5821 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5881 ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     5941 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     6001 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     6061 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     6121 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     6181 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     6241 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     6301 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6361 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6421 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6481 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6541 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6601 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6661 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6721 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6781 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6841 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6901 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6961 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     7021 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     7081 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     7141 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     7201 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     7261 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     7321 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7381 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7441 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7501 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7561 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7621 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7681 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7741 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7801 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7861 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7921 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7981 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     8041 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     8101 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     8161 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     8221 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     8281 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     8341 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     8401 caatttcaca caggaaacag ctatgacatg attacgaatt aa
//
