LOCUS       sequence-197628-        6112 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human Pim1 kinase domain in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6112)
  AUTHORS   Lee SJ, Han BG, Cho JW, Choi JS, Lee J, Song HJ, Koh JS, Lee BI
  TITLE     Crystal structure of pim1 kinase in complex with a 
            pyrido[4,3-d]pyrimidine derivative suggests a unique binding mode.
  JOURNAL   PLoS One. 2013 Jul 31;8(7):e70358. doi: 
            10.1371/journal.pone.0070358. Print 2013.
  PUBMED    23936194
REFERENCE   2  (bases 1 to 6112)
  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: "PLoS One.";
            date: "2013-07-31"; volume: "8(7):e70358. doi"; pages: " 
            10.1371/journal.pone.0070358. Print 2013"
COMMENT     Sequence Label: pHis-hPim1
FEATURES             Location/Qualifiers
     source          1..6112
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..92
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     180..199
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        180..198
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    199..223
                     /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)."
     CDS             1136..1159
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(1222..1240)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1236..1283
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1320..1775
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1407..1426)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1617..1638
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1867..2682)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1867..2682)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2590..2609
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2674..2693
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2804..3392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3293..3312
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3546..3563
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3578..3717
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3643..3662)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3803..3825
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3819..4010)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4819..5901)
                     /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     5863..5882
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5902..5979)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
       61 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
      121 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc ccgcgaaatt
      181 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag atttaaatta
      241 actttaagaa ggagatatac atatgaagga gaaggagccc ctggagtcgc agtaccaggt
      301 gggcccgcta ctgggcagcg gcggcttcgg ctcggtctac tcaggcatcc gcgtctccga
      361 caacttgccg gtggccatca aacacgtgga gaaggaccgg atttccgact ggggagagct
      421 gcctaatggc actcgagtgc ccatggaagt ggtcctgctg aagaaggtga gctcgggttt
      481 ctccggcgtc attaggctcc tggactggtt cgagaggccc gacagtttcg tcctgatcct
      541 ggagaggccc gagccggtgc aagatctctt cgacttcatc acggaaaggg gagccctgca
      601 agaggagctg gcccgcagct tcttctggca ggtgctggag gccgtgcggc actgccacaa
      661 ctgcggggtg ctccaccgcg acatcaagga cgaaaacatc cttatcgacc tcaatcgcgg
      721 cgagctcaag ctcatcgact tcgggtcggg ggcgctgctc aaggacaccg tctacacgga
      781 cttcgatggg acccgagtgt atagccctcc agagtggatc cgctaccatc gctaccatgg
      841 caggtcggcg gcagtctggt ccctggggat cctgctgtat gatatggtgt gtggagatat
      901 tcctttcgag catgacgaag agatcatcag gggccaggtt ttcttcaggc agagggtctc
      961 ttcagaatgt cagcatctca ttagatggtg cttggccctg agaccatcag ataggccaac
     1021 cttcgaagaa atccagaacc atccatggat gcaagatgtt ctcctgcccc aggaaactgc
     1081 tgagatccac ctccacagcc tgtcgccggg gcccagcaaa gcggccgcac tcgagcacca
     1141 ccaccaccac caccaccact aattgattaa tacctaggct gctaaacaaa gcccgaaagg
     1201 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     1261 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga
     1321 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1381 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1441 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1501 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1561 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1621 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1681 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1741 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     1801 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat
     1861 taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     1921 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     1981 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     2041 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     2101 gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac
     2161 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     2221 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     2281 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     2341 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     2401 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     2461 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     2521 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     2581 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     2641 tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     2701 gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt gagttttcgt
     2761 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2821 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2881 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2941 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3001 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3061 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3121 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3181 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3241 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3301 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3361 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3421 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3481 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3541 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     3601 cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc
     3661 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     3721 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3781 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     3841 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
     3901 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     3961 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     4021 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     4081 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     4141 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
     4201 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
     4261 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
     4321 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
     4381 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
     4441 gtcctcaacg acaggagcac gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg
     4501 gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg
     4561 tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg
     4621 tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag
     4681 aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact
     4741 gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt
     4801 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4861 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     4921 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     4981 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     5041 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     5101 gtccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     5161 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     5221 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     5281 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     5341 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     5401 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     5461 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     5521 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     5581 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     5641 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     5701 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     5761 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     5821 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     5881 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     5941 tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct
     6001 tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg
     6061 ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc ac
//
