LOCUS       sequence-439513-        5797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of FLAG-tagged human CAMK2D.
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
FEATURES             Location/Qualifiers
     source          1..5797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     170..189
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        303..407
                     /gene="bla"
                     /label=AmpR promoter
     promoter        409..766
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        409..600
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(432..452)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      617..752
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             801..1595
                     /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             801..1595
                     /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(855..874)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1465..1484
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1783..1802)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1827..1874
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2203..2791
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2692..2711
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2966..3269
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3270..3473
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3424..3444
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3519..3537
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      3572..3581
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3581..3604
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5197..5216)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5198..5216)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    5490..5611
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5527..5546)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5581..5600
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(join(5618..5797,1..276))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5755..5776)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
       61 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      121 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      181 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      241 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
      301 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      361 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagaatcct gaggcggaaa
      421 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
      481 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
      541 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
      601 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
      661 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
      721 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
      781 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
      841 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
      901 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
      961 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     1021 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     1081 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     1141 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     1201 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     1261 atcaggatga tctggacgaa gaacatcagg ggctcgcgcc agccgaactg ttcgccaggc
     1321 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     1381 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     1441 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gaacttggcg
     1501 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     1561 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     1621 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     1681 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     1741 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     1801 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     1861 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     1921 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     1981 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     2041 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     2101 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2161 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2221 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2281 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2341 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2401 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2461 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2521 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2581 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2641 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2701 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2761 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2821 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2881 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
     2941 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     3001 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     3061 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     3121 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     3181 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     3241 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     3301 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     3361 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     3421 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
     3481 tgaaccgtca gatccgctag cgattacgcc aagctcgaaa ttaaccctca ctaaagggaa
     3541 caaaagctgg agctccaccg cggtggcggc cgccaccatg gattacaagg atgacgacga
     3601 taagagcccg ggcggatccc ccgggctgca ggaattcatg gcttcgacca caacctgcac
     3661 caggttcacg gacgagtatc agcttttcga ggagcttgga aagggggcat tctcagtggt
     3721 gagaagatgt atgaaaattc ctactggaca agaatatgct gccaaaatta tcaacaccaa
     3781 aaagctttct gctagggatc atcagaaact agaaagagaa gctagaatct gccgtctttt
     3841 gaagcaccct aatattgtgc gacttcatga tagcatatca gaagagggct ttcactactt
     3901 ggtgtttgat ttagttactg gaggtgaact gtttgaagac atagtggcaa gagaatacta
     3961 cagtgaagct gatgccagtc attgtataca gcagattcta gaaagtgtta atcattgtca
     4021 cctaaatggc atagttcaca gggacctgaa gcctgagaat ttgcttttag ctagcaaatc
     4081 caagggagca gctgtgaaat tggcagactt tggcttagcc atagaagttc aaggggacca
     4141 gcaggcgtgg tttggttttg ctggcacacc tggatatctt tctccagaag ttttacgtaa
     4201 agatccttat ggaaagccag tggatatgtg ggcatgtggt gtcattctct atattctact
     4261 tgtggggtat ccacccttct gggatgaaga ccaacacaga ctctatcagc agatcaaggc
     4321 tggagcttat gattttccat caccagaatg ggacacggtg actcctgaag ccaaagacct
     4381 catcaataaa atgcttacta tcaaccctgc caaacgcatc acagcctcag aggcactgaa
     4441 gcacccatgg atctgtcaac gttctactgt tgcttccatg atgcacagac aggagactgt
     4501 agactgcttg aagaaattta atgctagaag aaaactaaag ggtgccatct tgacaactat
     4561 gctggctaca aggaatttct cagcagccaa gagtttgttg aagaaaccag atggagtaaa
     4621 ggagtcaact gagagttcaa atacaacaat tgaggatgaa gatgtgaaag cacgaaagca
     4681 agagattatc aaagtcactg aacaactgat cgaagctatc aacaatgggg actttgaagc
     4741 ctacacaaaa atctgtgacc caggccttac tgcttttgaa cctgaagctt tgggtaattt
     4801 agtggaaggg atggattttc accgattcta ctttgaaaat gctttgtcca aaagcaataa
     4861 accaatccac actattattc taaaccctca tgtacatctg gtaggggatg atgccgcctg
     4921 catagcatat attaggctca cacagtacat ggatggcagt ggaatgccaa agacaatgca
     4981 gtcagaagag actcgtgtgt ggcaccgccg ggatggaaag tggcagaatg ttcattttca
     5041 tcgctcgggg tcaccaacag tacccatcaa gccaccctgt attccaaatg ggaaagaaaa
     5101 cttctcagga ggcacctctt tgtggcaaaa catctaactc gagggggggc ccggtacctt
     5161 aattaattaa ggtaccaggt aagtgtaccc aattcgccct atagtgagtc gtattacaat
     5221 tcactcgatc gcccttccca acagttgcgc agcctgaatg gcgaatggag atccaatttt
     5281 taagtgtata atgtgttaaa ctactgattc taattgtttg tgtattttag attcacagtc
     5341 ccaaggctca tttcaggccc ctcagtcctc acagtctgtt catgatcata atcagccata
     5401 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     5461 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     5521 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5581 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
     5641 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     5701 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     5761 tggaacaaga gtccactatt aaagaacgtg gactcca
//
