LOCUS       sequence-439514-        5818 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of FLAG-tagged human CDC20.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5818)
  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 5818)
  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-CDC20
FEATURES             Location/Qualifiers
     source          1..5818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(799..822)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      822..831
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(866..884)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(930..1133)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(959..979)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1134..1437
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1612..2200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1692..1711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2529..2576
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2601..2620
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2808..3602)
                     /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(2808..3602)
                     /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(2919..2938)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3529..3548
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3637..3994)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3651..3786
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3803..3994
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     3951..3971
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(3996..4100)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4127..4582
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4214..4233)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4424..4445
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4589..4710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4600..4619)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4654..4673
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4984..5003
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4984..5002
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     5058..5074
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5059..5075
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cacatcccat agctgcacct cagcactgct ggtgcccaca gccaagtagt tgccctcttt
       61 gatccaggcc acagaggata tatattcccc aggctgctcc atttgcaaaa gctgcaggat
      121 gtcaccagag cttgcactcc acaggtacac actgttgtcc agtgccacgg ccagtacatt
      181 cccagaactc caatccacaa ggttcaggta atagtcattt cggatttcag gcgcatccag
      241 gatacggtct ggcagggaag gaatgtaacg gcaggtcttc cggctggagc caggagtggc
      301 cttttggctg tagagtactt tcagtctgtt ctgataaccc tctggcgcat tttgtggttt
      361 tccactgagc cgaaggatct tggcttcctc tacatcaaaa ccgttcaggt tcaaagccca
      421 ggctttctga tgttccttct tggtgggcgt ctggctgttt tcaggctggt tctccttgct
      481 caggaggaag ctggccacct ccatctgggc agcactgcga tgggggatat agcggtcacc
      541 gccaggtttg ctaggagtgg tctgaacctt ggaactggat ttgccaggag ttcggcccgg
      601 agtcctgccg gcgctgtggg atcggttggc ggcccgcatg ggtgaggggg ccgggcctgc
      661 ggcttccttg gctttgcgct gccagcgcgc agggggtgca ttggggatgg gtgcatccag
      721 ctgaagcagc gagtgcaggt cactctcgaa cgcgaactgt gccatgaatt cctgcagccc
      781 gggggatccg cccgggctct tatcgtcgtc atccttgtaa tccatggtgg cggccgccac
      841 cgcggtggag ctccagcttt tgttcccttt agtgagggtt aatttcgagc ttggcgtaat
      901 cgctagcgga tctgacggtt cactaaacca gctctgctta tatagacctc ccaccgtaca
      961 cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt
     1021 tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc
     1081 cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta
     1141 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac
     1201 tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc
     1261 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
     1321 cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat
     1381 gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg
     1441 aactccatat atgggctatg aactaatgac cccgtaattg attactatta ataactaatg
     1501 catggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1561 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1621 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1681 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1741 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1801 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1861 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1921 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1981 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2041 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2101 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2161 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2221 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2281 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2341 aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag
     2401 ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta
     2461 ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc
     2521 gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat
     2581 agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg
     2641 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     2701 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     2761 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     2821 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     2881 aggaagcggt cagcccattc gccgccaagt tcttcagcaa tatcacgggt agccaacgct
     2941 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     3001 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     3061 ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgt
     3121 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     3181 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     3241 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     3301 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     3361 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct
     3421 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     3481 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     3541 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     3601 atgcgaaacg atcctcatcc tgtctcttga tcgatctttg caaaagccta ggcctccaaa
     3661 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     3721 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     3781 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     3841 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     3901 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     3961 acacacattc cacagctggt tctttccgcc tcaggattct tcctttttca atattattga
     4021 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4081 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     4141 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4201 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4261 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4321 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4381 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4441 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4501 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4561 aaaatattaa cgcttacaat ttacgcgtta agatacattg atgagtttgg acaaaccaca
     4621 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     4681 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     4741 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     4801 atggctgatt atgatcatga acagactgtg aggactgagg ggcctgaaat gagccttggg
     4861 actgtgaatc taaaatacac aaacaattag aatcagtagt ttaacacatt atacacttaa
     4921 aaattggatc tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcgagtgaa
     4981 ttgtaatacg actcactata gggcgaattg ggtacactta cctggtacct taattaatta
     5041 aggtaccggg ccccccctcg aggtcgacgg tatcgataag ctttcagcgg atgccttggt
     5101 ggatgaggct gcttttggct gcactggcct tctcccgctc ccgccgccgc gcagggtcca
     5161 actcaaaaca gcgccatagc ctcagggtct catctgctgc tgcggatgcc actgtggccc
     5221 catctgggct catggtcaga ctcaggaccc gggatgtgtg acctttgagt tcagccacct
     5281 tggccatggt tgggtacttc caaataacta gctggttctg tgcaaagcca tggcctgaga
     5341 tgagctcctt gtaatgggga gaccagagga tggagcacac ctgggaatgg gcatccacgg
     5401 cactcagaca ggccccagag cacacattcc agatgcgaat gtgtcgatca ctggtgcccc
     5461 ctcctgttgc caggacattg gactgccagg gacaccatgc tacggccttg acagcccctt
     5521 gatgctgggt gaatgtctgc agaggaaccc agccaccctc tccaggagca ctaggccaca
     5581 cattgaccaa gttatcatta ccaccactgg ccaaatgtcg tccatctggg gcccagcgca
     5641 gcccacacac ttcctggctg tggccactca gtgtggccac atggtgttct gctacccgaa
     5701 catcatggtg gtggatgtgg ccagaacgtg aaccactgga caggatatag ctgttccagc
     5761 ttagggagcc cactcgggca gagtgactgg tcatatttcg aagccgtttc tgctgctg
//
