LOCUS       sequence-442615-        5774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of HA-tagged human p31(comet) (S102A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5774)
  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 5774)
  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: pCMV4-HA-p31(comet)-S102A
FEATURES             Location/Qualifiers
     source          1..5774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     254..273
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     507..524
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    641..662
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(677..682,683..700,701..707)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    715..731
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     720..742
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     739..755
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     739..755
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     769..788
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        769..787
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          864..960
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     promoter        complement(999..1328)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1013..1148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1313..1333
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    1357..1979
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1761..1780
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(2833..2922)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     promoter        complement(3020..3223)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3049..3069)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3224..3603
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3764..3781)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3764..3780)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3773..3795)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3921..4376)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4058..4079)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4270..4289
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4403..4507
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4508..4576,4577..5368)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4726..4745)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5539..5774,1..353)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
       61 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      121 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      181 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      241 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      301 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      361 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      421 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      481 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
      541 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
      601 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
      661 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
      721 gcggataaca atttcacaca ggaaacagct atgacatgat tacgaattta atacgactca
      781 ctatagggaa ttcgagctcg gtacgcggcc ctagagtcga cctgcagcag cttttagagc
      841 agaagtaaca cttccgtaca ggcctagaag taaaggcaac atccactgag gagcagttct
      901 ttgatttgca ccaccaccgg atccgggacc tgaaataaaa gacaaaaaga ctaaacttac
      961 cagttaactt tctggttttt cagttcctcg aggagctttt tgcaaaagcc taggcctcca
     1021 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca
     1081 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     1141 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     1201 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     1261 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     1321 tgacacacat tccacagcca agctggccgc gtacccaatt caacaggcat ctactgagtg
     1381 gacccaacgc atgagaggac agtgccaagc aagcaactca aatgtcccac cggttgggca
     1441 tggccaggta gcctatgctg tgtctggacg tcctcctgct ggtatagtta ttttaaaatc
     1501 agaaggacag ggaagggagc agtggttcac gcctgtaatc ccagcaattt gggaggccaa
     1561 ggtgggtaga tcacctgaga ttaggagttg gagaccagcc tggccaatat ggtgaaaccc
     1621 cgtctctacc aaaaaaacaa aaattagctg agcctggtca tgcatgcctg gaatcccaac
     1681 aactcgggag gctgaggcag gagaatcgct tgaacccagg aggcggagat tgcagtgagc
     1741 caagattgtg ccactgcact ccagcttggt tcccaataga ccccgcaggc cctacaggtt
     1801 gtcttcccaa cttgcccctt gctccatacc acccccctcc accccataat attatagaag
     1861 gacacctagt cagacaaaat gatgcaactt aattttatta ggacaaggct ggtgggcact
     1921 ggagtggcaa cttccagggc caggagaggc actggggagg ggtcacaggg atgccacccg
     1981 ggatcctcta gactcgcgga agcctttaaa tgtcactggt gcctggaacc aaatgtagtc
     2041 ttcccaagcc gtggttcgga tggaaggtct gccacatgac agggtcacag tcagtttatg
     2101 gccccggctg ggcactcgat agttgagctt gggtcgaaac caatcttctc cacagttgcg
     2161 gtgtccctgt gccatgacga cggtgcccat gagtggagga gcctgaagct cgctaaaggc
     2221 atcagccatg aatatggctc ggaagagacg gcgcaaacaa gctgctgtgc tcaggctctg
     2281 gtccacgctg tagggggcca gcagagacaa gtcgagttca tagaactcct tggggcttag
     2341 ggcattgccc ccaaggagaa tcagcactcg cggtactagt gtccgtgcaa agaagtcctc
     2401 caggtggctg aggacactct ccagttctgc cagggcttgt tggcatttcc tgctgctcac
     2461 ctcagtggtg gcccgaggtt tcttcttcag catctcctct gcctggggag caggttttcg
     2521 gtaaaagtgc ttaagctgtt catagggcag agggagctgc tggcgttgat acatgatatg
     2581 ctttagaagt tcacaagtaa actgacagca gccttcctgg ctcacaggcc caggaaacac
     2641 cactggtacc atgcagtctc ttgggcaaaa ggcctccgaa gcgttgagag gttcctgcgt
     2701 agagcttgtc tcaagtagtt ctatctggga ggcgtgagtt tcttcggact tctcatacca
     2761 ctccaaatca gggactgcgg ctgaggacag aacctccgcc tccggcgccg ccataagctt
     2821 atcgatctga gcagcgtaat ctggaacgtc atatggatag gatcctgcat agtccgggac
     2881 gtcataggga tagcccgcat agtcaggaac atcgtatggg taaaagatat ccatggtacg
     2941 cgtggtacca gatctgaatt cgatggaagt atttgaaaga aaattaaaaa taaaaacaat
     3001 tctgacggtt cactaaacga gctctgctta tatagacctc ccaccgtaca cgcctaccgc
     3061 ccatttgcgt caacggggcg gggttattac gacattttgg aaagtcccgt tgattttggt
     3121 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa
     3181 accgctatcc acgcccattg gtgtactgcc aaaaccgcat caccatggta atagcgatga
     3241 ctaatacgta gatgtactgc caagtaggaa agtcccgtaa ggtcatgtac tgggcataat
     3301 gccaggcggg ccatttaccg tcattgacgt caataggggg cggacttggc atatgataca
     3361 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     3421 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     3481 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     3541 atgggctatg aactaatgac cccgtaattg attactatta ataactagtc aataatcaat
     3601 gtcaacatgg cggtcatatt ggacatgagc caatataaat gtacatatta tgatatagat
     3661 acaacgtatg caatggccaa tagccaatat tgatttatgc tatataacca atgactaata
     3721 tggctaattg ccaatattga ttcaatgtat agatcagctt ggcactggcc gtcgttttac
     3781 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     3841 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     3901 gcagcctgaa tggcgaatgg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     3961 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     4021 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     4081 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4141 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4201 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     4261 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     4321 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     4381 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4441 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4501 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4561 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4621 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4681 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4741 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4801 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4861 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4921 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4981 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5041 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5101 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5161 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5221 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5281 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5341 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5401 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5461 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5521 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5581 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5641 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5701 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5761 cctgttacca gtgg
//
