LOCUS       sequence-439897-        5143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of FLAG-tagged human p31(comet) (Q83A/F191A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5143)
  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 5143)
  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-p31(comet)-QF
FEATURES             Location/Qualifiers
     source          1..5143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(188..776)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(268..287)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1105..1152
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1177..1196
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1384..2178)
                     /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(1384..2178)
                     /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(1495..1514)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2105..2124
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2213..2570)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2227..2362
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2379..2570
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2527..2547
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(2572..2676)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2703..3158
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2790..2809)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3000..3021
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3165..3286
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3176..3195)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3230..3249
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3560..3579
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3560..3578
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3634..3650
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3635..3651
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4518..4541)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      4541..4550
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4585..4603)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(4649..4852)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4678..4698)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(4853..5143,1..13)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
       61 ctattaataa ctaatgcatg gcggtaatac ggttatccac agaatcaggg gataacgcag
      121 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      181 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      241 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      301 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      361 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      421 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
      481 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
      541 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
      601 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
      661 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
      721 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
      781 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
      841 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
      901 gttttaaatc aatctaaagt atatatgagt aacctgaggc tatggcaggg cctgccgccc
      961 cgacgttggc tgcgagccct gggccttcac ccgaacttgg ggggtggggt ggggaaaagg
     1021 aagaaacgcg ggcgtattgg ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc
     1081 cctgggaccg aaccccgcgt ttatgaacaa acgacccaac accgtgcgtt ttattctgtc
     1141 tttttattgc cgtcatagcg cgggttcctt ccggtattgt ctccttccgt gtttcagtta
     1201 gcctccccct agggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc
     1261 agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat
     1321 cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt catttcgaac cccagagtcc
     1381 cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc
     1441 gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagttctt cagcaatatc
     1501 acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat
     1561 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt
     1621 cacgacgaga tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg
     1681 cgcgagcccc tgatgttctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
     1741 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
     1801 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
     1861 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
     1921 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
     1981 ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
     2041 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
     2101 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
     2161 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa
     2221 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
     2281 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
     2341 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
     2401 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
     2461 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     2521 ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag gattcttcct
     2581 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     2641 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     2701 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2761 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2821 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2881 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2941 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3001 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3061 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3121 taacgcgaat tttaacaaaa tattaacgct tacaatttac gcgttaagat acattgatga
     3181 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3241 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3301 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     3361 cctctacaaa tgtggtatgg ctgattatga tcatgaacag actgtgagga ctgaggggcc
     3421 tgaaatgagc cttgggactg tgaatctaaa atacacaaac aattagaatc agtagtttaa
     3481 cacattatac acttaaaaat tggatctcca ttcgccattc aggctgcgca actgttggga
     3541 agggcgatcg agtgaattgt aatacgactc actatagggc gaattgggta cacttacctg
     3601 gtaccttaat taattaaggt accgggcccc ccctcgaggt cgacggtatc gataagcttt
     3661 cactcgcgga agcctttaaa tgtcactggt gcctggaacc aaatgtagtc ttcccaagcc
     3721 gtggttcgga tggaaggtct gccacatgac agggtcacag tcagtttatg gccccggctg
     3781 ggcactcgat agttgagctt gggtcgaaac caatcttctc cacagttgcg gtgtccctgt
     3841 gccatgacga cggtgcccat gagtggagga gcctgaagct cgctaaaggc atcagccatg
     3901 aatatggctc gggcgagacg gcgcaaacaa gctgctgtgc tcaggctctg gtccacgctg
     3961 tagggggcca gcagagacaa gtcgagttca tagaactcct tggggcttag ggcattgccc
     4021 ccaaggagaa tcagcactcg cggtactagt gtccgtgcaa agaagtcctc caggtggctg
     4081 aggacactct ccagttctgc cagggcttgt tggcatttcc tgctgctcac ctcagtggtg
     4141 gcccgaggtt tcttcttcag catctcctct gcctggggag aaggttttcg gtaaaagtgc
     4201 ttaagctgtt catagggcag agggagctgc tggcgtgcat acatgatatg ctttagaagt
     4261 tcacaagtaa actgacagca gccttcctgg ctcacaggcc caggaaacac cactggtacc
     4321 atgcagtctc ttgggcaaaa ggcctccgaa gcgttgagag gttcctgcgt agagcttgtc
     4381 tcaagtagtt ctatctggga ggcgtgagtt tcttcggact tctcatacca ctccaaatca
     4441 gggactgcgg ctgaggacag aacctccgcc tccggcgccg ccatgaattc ctgcagcccg
     4501 ggggatccgc ccgggctctt atcgtcgtca tccttgtaat ccatggtggc ggccgccacc
     4561 gcggtggagc tccagctttt gttcccttta gtgagggtta atttcgagct tggcgtaatc
     4621 gctagcggat ctgacggttc actaaaccag ctctgcttat atagacctcc caccgtacac
     4681 gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt
     4741 gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc
     4801 gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa
     4861 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
     4921 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
     4981 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     5041 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     5101 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgt
//
