LOCUS       sequence-439511-        4933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of FLAG-tagged human MAD2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4933)
  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 4933)
  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-MAD2
FEATURES             Location/Qualifiers
     source          1..4933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(147..170)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      170..179
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(214..232)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(278..481)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(307..327)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        482..785
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(960..1548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1040..1059)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1877..1924
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1949..1968
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2156..2950)
                     /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(2156..2950)
                     /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(2267..2286)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2877..2896
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2985..3342)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2999..3134
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3151..3342
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     3299..3319
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(3344..3448)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3475..3930
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3562..3581)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3772..3793
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3937..4058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3948..3967)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4002..4021
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4332..4351
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4332..4350
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4406..4422
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4407..4423
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tatatgccac gctgatataa aatgctgttg atgccgaatg agaagaactc ggccacgatt
       61 tcggcgctcc cgcgcagggt gattccctgc tcccgggaga gctgcagcgc catgaattcc
      121 tgcagcccgg gggatccgcc cgggctctta tcgtcgtcat ccttgtaatc catggtggcg
      181 gccgccaccg cggtggagct ccagcttttg ttccctttag tgagggttaa tttcgagctt
      241 ggcgtaatcg ctagcggatc tgacggttca ctaaaccagc tctgcttata tagacctccc
      301 accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa
      361 agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg
      421 gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca
      481 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
      541 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
      601 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
      661 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
      721 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
      781 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
      841 aactaatgca tggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
      901 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
      961 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1021 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1081 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1141 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1201 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1261 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1321 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1381 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     1441 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     1501 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     1561 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     1621 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     1681 tcaatctaaa gtatatatga gtaacctgag gctatggcag ggcctgccgc cccgacgttg
     1741 gctgcgagcc ctgggccttc acccgaactt ggggggtggg gtggggaaaa ggaagaaacg
     1801 cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac
     1861 cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg ttttattctg tctttttatt
     1921 gccgtcatag cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt tagcctcccc
     1981 ctagggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg
     2041 tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct
     2101 cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga
     2161 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
     2221 aaagcacgag gaagcggtca gcccattcgc cgccaagttc ttcagcaata tcacgggtag
     2281 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag
     2341 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
     2401 gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc
     2461 cctgatgttc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     2521 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     2581 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     2641 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     2701 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     2761 cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     2821 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     2881 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     2941 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca aaagcctagg
     3001 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     3061 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     3121 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
     3181 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
     3241 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     3301 cctaactgac acacattcca cagctggttc tttccgcctc aggattcttc ctttttcaat
     3361 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     3421 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc
     3481 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3541 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3601 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3661 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3721 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3781 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3841 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3901 attttaacaa aatattaacg cttacaattt acgcgttaag atacattgat gagtttggac
     3961 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     4021 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     4081 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     4141 aatgtggtat ggctgattat gatcatgaac agactgtgag gactgagggg cctgaaatga
     4201 gccttgggac tgtgaatcta aaatacacaa acaattagaa tcagtagttt aacacattat
     4261 acacttaaaa attggatctc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     4321 cgagtgaatt gtaatacgac tcactatagg gcgaattggg tacacttacc tggtacctta
     4381 attaattaag gtaccgggcc ccccctcgag gtcgacggta tcgataagct tgtcattgac
     4441 aggaattttg taggccacca tgctatttac tttgtggatt gtagtagtaa atgaacgaag
     4501 gcggacttcc tcagaattgg taataaactg tggtcccgac tcttcccatt tttcaggtac
     4561 aaccaaatct ttgtctgtat aaatcagcag atcaaatgaa caagaaactt ccaacagtgg
     4621 cagaaatgtc accgtagctg tgatctgtct gatcactgaa cggatttcat cctggatagc
     4681 tttctgagac ttttctctgg gtgcactgtc atcttttgca gtcttgtcac actcaatatc
     4741 aaactgccat ctttccagga cctcaccact ttcaatattt gagataacta caaccagttt
     4801 ctgaactgaa cacttgtata accaatcttt cagttgttcc accacattat ttaggtattt
     4861 tatgagctca agatcagtag ttacaagcaa ggtgagtccg tatttctgca ctcgagtaaa
     4921 ggtttcagat gga
//
