LOCUS       sequence-439510-        5797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of FLAG-tagged human CAMK2D (T287A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5797)
  AUTHORS   Chuah YH, Tay EXY, Grinchuk OV, Yoon J, Feng J, Kannan S, Robert M, 
            Jakhar R, Liang Y, Lee BWL, Wang LC, Lim YT, Zhao T, Sobota RM, Lu 
            G, Low BC, Crasta KC, Verma CS, Lin Z, Ong DST
  TITLE     CAMK2D serves as a molecular scaffold for RNF8-MAD2 complex to 
            induce mitotic checkpoint in glioma.
  JOURNAL   Cell Death Differ. 2023 Aug;30(8):1973-1987. doi: 
            10.1038/s41418-023-01192-3. Epub 2023 Jul 19.
  PUBMED    37468549
REFERENCE   2  (bases 1 to 5797)
  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: 
            "10.1038/s41418-023-01192-3"; journalName: "Cell Death Differ"; 
            date: "2023-08"; volume: "30"; issue: "8"; pages: "1973-1987"
COMMENT     Sequence Label: pCMV-FLAG-CAMK2D-T287A
FEATURES             Location/Qualifiers
     source          1..5797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..46
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(71..175)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      202..657
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(289..308)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     499..520
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    664..785
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(675..694)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     729..748
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1059..1078
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1059..1077
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2671..2694)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2694..2703
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2738..2756)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(2802..3005)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2831..2851)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3006..3309
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3484..4072)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3564..3583)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4401..4448
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4473..4492
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4680..5474)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4680..5474)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4791..4810)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5401..5420
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5509..5797,1..69))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5523..5658
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        join(5675..5797,1..69)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
ORIGIN
        1 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt
       61 ccgcctcagg attcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
      121 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
      181 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
      241 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
      301 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
      361 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
      421 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
      481 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
      541 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
      601 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttacg
      661 cgttaagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
      721 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
      781 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
      841 ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat catgaacaga
      901 ctgtgaggac tgaggggcct gaaatgagcc ttgggactgt gaatctaaaa tacacaaaca
      961 attagaatca gtagtttaac acattataca cttaaaaatt ggatctccat tcgccattca
     1021 ggctgcgcaa ctgttgggaa gggcgatcga gtgaattgta atacgactca ctatagggcg
     1081 aattgggtac acttacctgg taccttaatt aattaaggta ccgggccccc cctcgagtta
     1141 gatgttttgc cacaaagagg tgcctcctga gaagttttct ttcccatttg gaatacaggg
     1201 tggcttgatg ggtactgttg gtgaccccga gcgatgaaaa tgaacattct gccactttcc
     1261 atcccggcgg tgccacacac gagtctcttc tgactgcatt gtctttggca ttccactgcc
     1321 atccatgtac tgtgtgagcc taatatatgc tatgcaggcg gcatcatccc ctaccagatg
     1381 tacatgaggg tttagaataa tagtgtggat tggtttattg cttttggaca aagcattttc
     1441 aaagtagaat cggtgaaaat ccatcccttc cactaaatta cccaaagctt caggttcaaa
     1501 agcagtaagg cctgggtcac agatttttgt gtaggcttca aagtccccat tgttgatagc
     1561 ttcgatcagt tgttcagtga ctttgataat ctcttgcttt cgtgctttca catcttcatc
     1621 ctcaattgtt gtatttgaac tctcagttga ctcctttact ccatctggtt tcttcaacaa
     1681 actcttggct gctgagaaat tccttgtagc cagcatagtt gtcaagatgg caccctttag
     1741 ttttcttcta gcattaaatt tcttcaagca gtctacagcc tcctgtctgt gcatcatgga
     1801 agcaacagta gaacgttgac agatccatgg gtgcttcagt gcctctgagg ctgtgatgcg
     1861 tttggcaggg ttgatagtaa gcattttatt gatgaggtct ttggcttcag gagtcaccgt
     1921 gtcccattct ggtgatggaa aatcataagc tccagccttg atctgctgat agagtctgtg
     1981 ttggtcttca tcccagaagg gtggataccc cacaagtaga atatagagaa tgacaccaca
     2041 tgcccacata tccactggct ttccataagg atctttacgt aaaacttctg gagaaagata
     2101 tccaggtgtg ccagcaaaac caaaccacgc ctgctggtcc ccttgaactt ctatggctaa
     2161 gccaaagtct gccaatttca cagctgctcc cttggatttg ctagctaaaa gcaaattctc
     2221 aggcttcagg tccctgtgaa ctatgccatt taggtgacaa tgattaacac tttctagaat
     2281 ctgctgtata caatgactgg catcagcttc actgtagtat tctcttgcca ctatgtcttc
     2341 aaacagttca cctccagtaa ctaaatcaaa caccaagtag tgaaagccct cttctgatat
     2401 gctatcatga agtcgcacaa tattagggtg cttcaaaaga cggcagattc tagcttctct
     2461 ttctagtttc tgatgatccc tagcagaaag ctttttggtg ttgataattt tggcagcata
     2521 ttcttgtcca gtaggaattt tcatacatct tctcaccact gagaatgccc cctttccaag
     2581 ctcctcgaaa agctgatact cgtccgtgaa cctggtgcag gttgtggtcg aagccatgaa
     2641 ttcctgcagc ccgggggatc cgcccgggct cttatcgtcg tcatccttgt aatccatggt
     2701 ggcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg ttaatttcga
     2761 gcttggcgta atcgctagcg gatctgacgg ttcactaaac cagctctgct tatatagacc
     2821 tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt
     2881 ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga
     2941 cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc
     3001 atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat
     3061 aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg
     3121 ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac
     3181 tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt
     3241 attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt
     3301 tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat
     3361 taataactaa tgcatggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3421 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3481 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3541 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3601 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3661 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3721 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3781 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3841 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3901 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3961 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4021 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4081 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4141 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4201 taaatcaatc taaagtatat atgagtaacc tgaggctatg gcagggcctg ccgccccgac
     4261 gttggctgcg agccctgggc cttcacccga acttgggggg tggggtgggg aaaaggaaga
     4321 aacgcgggcg tattggcccc aatggggtct cggtggggta tcgacagagt gccagccctg
     4381 ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg tgcgttttat tctgtctttt
     4441 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
     4501 ccccctaggg tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc
     4561 ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa
     4621 atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct
     4681 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     4741 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gttcttcagc aatatcacgg
     4801 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat
     4861 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
     4921 acgagatcct cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg
     4981 agcccctgat gttcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     5041 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     5101 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     5161 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     5221 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     5281 gctgcctcgt cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     5341 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     5401 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     5461 tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc
     5521 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     5581 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     5641 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     5701 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     5761 tgactaattg agatgcatgc tttgcatact tctgcct
//
