LOCUS       sequence-439898-        5797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of FLAG-tagged human CAMK2D (K43R/D136N).
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-ED
FEATURES             Location/Qualifiers
     source          1..5797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     466..485
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(784..803)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    828..875
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1204..1792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1693..1712
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1967..2270
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2271..2474
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2425..2445
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2520..2538
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      2573..2582
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2582..2605
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4198..4217)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4199..4217)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4491..4612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4528..4547)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4582..4601
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4619..5074)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4756..4777)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4968..4987
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5101..5205
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5207..5564
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5207..5398
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5230..5250)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5415..5550
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5599..5797,1..596)
                     /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             join(5599..5797,1..596)
                     /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(5653..5672)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
       61 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
      121 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
      181 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
      241 ggatggaagc cggtcttgtc gatcaggatg atctggacga agaacatcag gggctcgcgc
      301 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
      361 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
      421 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
      481 atattgctga agaacttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
      541 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
      601 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
      661 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
      721 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
      781 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
      841 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
      901 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
      961 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     1021 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     1081 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     1141 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1201 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1261 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1321 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     1381 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     1441 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     1501 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     1561 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     1621 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     1681 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     1741 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     1801 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     1861 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
     1921 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     1981 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     2041 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     2101 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     2161 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     2221 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     2281 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     2341 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     2401 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     2461 atataagcag agctggttta gtgaaccgtc agatccgcta gcgattacgc caagctcgaa
     2521 attaaccctc actaaaggga acaaaagctg gagctccacc gcggtggcgg ccgccaccat
     2581 ggattacaag gatgacgacg ataagagccc gggcggatcc cccgggctgc aggaattcat
     2641 ggcttcgacc acaacctgca ccaggttcac ggacgagtat cagcttttcg aggagcttgg
     2701 aaagggggca ttctcagtgg tgagaagatg tatgaaaatt cctactggac aagaatatgc
     2761 tgccagaatt atcaacacca aaaagctttc tgctagggat catcagaaac tagaaagaga
     2821 agctagaatc tgccgtcttt tgaagcaccc taatattgtg cgacttcatg atagcatatc
     2881 agaagagggc tttcactact tggtgtttga tttagttact ggaggtgaac tgtttgaaga
     2941 catagtggca agagaatact acagtgaagc tgatgccagt cattgtatac agcagattct
     3001 agaaagtgtt aatcattgtc acctaaatgg catagttcac aggaacctga agcctgagaa
     3061 tttgctttta gctagcaaat ccaagggagc agctgtgaaa ttggcagact ttggcttagc
     3121 catagaagtt caaggggacc agcaggcgtg gtttggtttt gctggcacac ctggatatct
     3181 ttctccagaa gttttacgta aagatcctta tggaaagcca gtggatatgt gggcatgtgg
     3241 tgtcattctc tatattctac ttgtggggta tccacccttc tgggatgaag accaacacag
     3301 actctatcag cagatcaagg ctggagctta tgattttcca tcaccagaat gggacacggt
     3361 gactcctgaa gccaaagacc tcatcaataa aatgcttact atcaaccctg ccaaacgcat
     3421 cacagcctca gaggcactga agcacccatg gatctgtcaa cgttctactg ttgcttccat
     3481 gatgcacaga caggagactg tagactgctt gaagaaattt aatgctagaa gaaaactaaa
     3541 gggtgccatc ttgacaacta tgctggctac aaggaatttc tcagcagcca agagtttgtt
     3601 gaagaaacca gatggagtaa aggagtcaac tgagagttca aatacaacaa ttgaggatga
     3661 agatgtgaaa gcacgaaagc aagagattat caaagtcact gaacaactga tcgaagctat
     3721 caacaatggg gactttgaag cctacacaaa aatctgtgac ccaggcctta ctgcttttga
     3781 acctgaagct ttgggtaatt tagtggaagg gatggatttt caccgattct actttgaaaa
     3841 tgctttgtcc aaaagcaata aaccaatcca cactattatt ctaaaccctc atgtacatct
     3901 ggtaggggat gatgccgcct gcatagcata tattaggctc acacagtaca tggatggcag
     3961 tggaatgcca aagacaatgc agtcagaaga gactcgtgtg tggcaccgcc gggatggaaa
     4021 gtggcagaat gttcattttc atcgctcggg gtcaccaaca gtacccatca agccaccctg
     4081 tattccaaat gggaaagaaa acttctcagg aggcacctct ttgtggcaaa acatctaact
     4141 cgaggggggg cccggtacct taattaatta aggtaccagg taagtgtacc caattcgccc
     4201 tatagtgagt cgtattacaa ttcactcgat cgcccttccc aacagttgcg cagcctgaat
     4261 ggcgaatgga gatccaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt
     4321 gtgtatttta gattcacagt cccaaggctc atttcaggcc cctcagtcct cacagtctgt
     4381 tcatgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     4441 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     4501 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4561 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     4621 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4681 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4741 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4801 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     4861 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     4921 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4981 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     5041 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     5101 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5161 caataaccct gataaatgct tcaataatat tgaaaaagga agaatcctga ggcggaaaga
     5221 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     5281 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     5341 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     5401 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5461 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     5521 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     5581 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5641 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5701 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5761 ggtgccctga atgaactgca agacgaggca gcgcggc
//
