LOCUS       sequence-215941-        8538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of SRPK2_Pkinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8538)
  AUTHORS   Teyra J, Kelil A, Jain S, Helmy M, Jajodia R, Hooda Y, Gu J, D'Cruz 
            AA, Nicholson SE, Min J, Sudol M, Kim PM, Bader GD, Sidhu SS
  TITLE     Large-scale survey and database of high affinity ligands for peptide
            recognition modules.
  JOURNAL   Mol Syst Biol. 2020 Dec;16(12):e9310. doi: 10.15252/msb.20199310.
  PUBMED    33438817
REFERENCE   2  (bases 1 to 8538)
  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 Syst 
            Biol."; date: "2020-12"; volume: "16(12):e9310. doi"; pages: " 
            10.15252/msb.20199310"
COMMENT     Sequence Label: pHH0103_SRPK2_Pkinase
FEATURES             Location/Qualifiers
     source          1..8538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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     42..64
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             138..155
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             171..824
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     782..804
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             831..854
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             867..884
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2733..2780
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4373..4828)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4510..4531)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4722..4741
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4870..4892)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4978..5118
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5033..5052
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5133..5150)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5304..5891)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6062..6853,6854..6922))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6685..6704
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6923..7027)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7095..7113
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7136..7213
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             7214..8296
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(7233..7252)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccataga
      121 ttatgaaaat cgaagaacac catcaccatc accattccag cggtaagctt atgtccccta
      181 tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc
      241 ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca
      301 aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta
      361 aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg
      421 gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat
      481 acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta
      541 gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa
      601 atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat
      661 acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg
      721 aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc
      781 agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat ctggaagttc
      841 tgttccaggg gcccctgtcc agcggtctgg ttccgcgtgg ttccggtacc gcggcccagc
      901 cggcctatca tgtcattcgc aaactgggct ggggtcactt ttcgaccgtg tggctgtgct
      961 gggatatgca gggcaaacgt ttcgttgcga tgaaagtggt taaaagcgcc caacattaca
     1021 ccgaaacggc actggatgaa attaaactgc tgaaatgtgt gcgcgaatct gacccgagtg
     1081 atccgaataa agatatggtc gtgcagctga ttgatgactt taaaatctct ggcatgaacg
     1141 gtattcacgt gtgcatggtt ttcgaagtcc tgggccatca cctgctgaaa tggattatca
     1201 aatcgaatta tcagggtctg ccggtgcgtt gtgttaaaag cattatccgc caggtgctgc
     1261 aaggcctgga ttacctgcat agtaaatgca aaattatcca caccgacatt aaaccggaaa
     1321 acatcctgat gtgtgtcgat gacgcttatg tgcgtcgcat ggcggccgaa gcgaccgaat
     1381 ggcagaaagc aggtgctccg ccgccgtctg gtagtgcagt ttctacggct ccgcagcaaa
     1441 aaccgattgg taaaatcagt aaaaacaaaa agaaaaaact gaagaaaaaa cagaaacgtc
     1501 aagcagaact gctggaaaaa cgcctgcagg aaattgaaga actggaacgt gaagcggaac
     1561 gcaaaattat cgaagaaaat atcaccagcg cagctccgtc taacgaccag gatggcgaat
     1621 attgcccgga agttaaactg aaaaccacgg gtctggaaga agcggccgaa gcagaaacgg
     1681 ctaaagacaa cggcgaagcc gaagatcagg aagaaaaaga agacgcagaa aaagaaaaca
     1741 tcgaaaaaga cgaagatgac gtcgatcaag aactggcgaa cattgatccg acctggatcg
     1801 aatccccgaa aacgaacggc catatcgaaa atggtccgtt ttcactggaa cagcaactgg
     1861 atgacgaaga tgacgatgaa gaagattgcc cgaacccgga agaatataat ctggatgaac
     1921 cgaacgccga atctgactat acctacagct ctagttacga acagtttaat ggcgaactgc
     1981 cgaacggtcg ccataaaatt ccggaaagtc aatttccgga attctccacc tcactgttct
     2041 cgggtagcct ggaaccggtt gcgtgtggct ccgtcctgtc agaaggttcg ccgctgacgg
     2101 aacaggaaga atcctcaccg agtcacgatc gttcccgtac cgtctcagct tcgagcacgg
     2161 gtgatctgcc gaaagcaaaa acccgtgcag ctgacctgct ggtgaatccg ctggacccgc
     2221 gtaacgccga taaaattcgc gtgaaaatcg cggatctggg taatgcctgc tgggttcata
     2281 aacactttac cgaagatatt cagacgcgtc aatatcgcag cattgaagtg ctgatcggcg
     2341 caggttactc caccccggct gatatctggt caacggcgtg tatggccttt gaactggcga
     2401 ccggcgatta cctgtttgaa ccgcattctg gtgaagacta cagtcgtgac gaagatcata
     2461 ttgcgcacat tatcgaactg ctgggctcca tcccgcgtca ttttgccctg tcgggtaaat
     2521 atagccgcga atttttcaac cgtcgcggcg aactgcgtca cattaccaaa ctgaaaccgt
     2581 ggagcctgtt cgatgttctg gtcgaaaaat acggttggcc gcatgaagat gcggcccagt
     2641 ttacggactt cctgatcccg atgctggaaa tggttccgga aaaacgtgcg agcgccggtg
     2701 aatgtctgcg ccacccggcg gccgcataat aaaccgatac aattaaaggc tccttttgga
     2761 gccttttttt tggagatttt caacgtgaaa aaattattat tcgcaattcc tttagttgtt
     2821 cctttctatt ctcactccgc tgaaactgtt gaaagttgtt tagcaaaacc ccatacagaa
     2881 aattcattta ctaacgtctg gaaagacgac aaaactttag atcgttacgc taactatgag
     2941 ggctgtctgt ggaatgctac aggcgttgta gtttgtactg gtgacgaaac tcagtgtcta
     3001 gctagagtgg cggtggctct ggttccggtg attttgatta tgaaaagatg gcaaacgcta
     3061 ataagggggc tatgaccgaa aatgccgatg aaaacgcgct acagtctgac gctaaaggca
     3121 aacttgattc tgtcgctact gattacggtg ctgctatcga tggtttcatt ggtgacgttt
     3181 ccggccttgc taatggtaat ggtgctactg gtgattttgc tggctctaat tcccaaatgg
     3241 ctcaagtcgg tgacggtgat aattcacctt taatgaataa tttccgtcaa tatttacctt
     3301 ccctccctca atcggttgaa tgtcgccctt ttgtctttgg cgctggtaaa ccatatgaat
     3361 tttctattga ttgtgacaaa ataaacttat tccgtggtgt ctttgcgttt cttttatatg
     3421 ttgccacctt tatgtatgta ttttctacgt ttgctaacat actgcgtaat aaggagtctt
     3481 aatcatgcca gttcttttgg ctagcgccgc cctatacctt gtctgcctcc ccgcgttgcg
     3541 tcgcggtgca tggagccggg ccacctcgac ctgaatggaa gccggcggca cctcgctaac
     3601 ggattcacca ctccaagaat tggagccaat caattcttgc ggagaactgt gaatgcgcaa
     3661 accaaccctt ggcagaacat atccatcgcg tccgccatct ccagcagccg cacgcggcgc
     3721 atctcgggca gcgttgggtc ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg
     3781 acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag
     3841 cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc
     3901 ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cagcgccctg caccattatg
     3961 ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac atctgtatta
     4021 acgaagcgct ggcattgacc ctgagtgatt tttctctggt cccgccgcat ccataccgcc
     4081 agttgtttac cctcacaacg ttccagtaac cgggcatgtt catcatcagt aacccgtatc
     4141 gtgagcatcc tctctcgttt catcggtatc attaccccca tgaacagaaa tcccccttac
     4201 acggaggcat cagtgaccaa acaggaaaaa accgccctta acatggcccg ctttatcaga
     4261 agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga acaggcagac
     4321 atctgtgaat cgcttcacga ccacgctgat gagctttacc gcaggatccg cgaaattgta
     4381 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4441 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4501 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4561 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4621 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4681 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4741 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4801 gcgcttaatg cgccgctaca gggcgcgtcg cggatcctgc ctcgcgcgtt tcggtgatga
     4861 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     4921 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     4981 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
     5041 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     5101 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5161 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5221 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5281 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5341 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccagggtttc
     5401 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5461 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5521 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5581 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5641 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5701 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     5761 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5821 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5881 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5941 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6001 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6061 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6121 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6181 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6241 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6301 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6361 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6421 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6481 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6541 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6601 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6661 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6721 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6781 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6841 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6901 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6961 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7021 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     7081 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttcccgacac
     7141 catcgaatgg tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt
     7201 cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc
     7261 ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga
     7321 aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact
     7381 ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc
     7441 gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt
     7501 gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc
     7561 gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt
     7621 ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat
     7681 caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc
     7741 attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct
     7801 gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg
     7861 ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg
     7921 catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc
     7981 cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac
     8041 cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct
     8101 ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa
     8161 tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac
     8221 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     8281 ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacaat
     8341 tctcatgttt gacagcttat catcgactgc acggtgcacc aatgcttctg gcgtcaggca
     8401 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
     8461 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
     8521 atattctgaa atgagctg
//
