LOCUS       sequence-442613-        5774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of HA-tagged human p31(comet).
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)
FEATURES             Location/Qualifiers
     source          1..5774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        216..419
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     370..390
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             517..606
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     polyA_signal    1460..2082
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1659..1678)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2106..2126)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2111..2440
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2291..2426
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          2479..2575
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     primer_bind     complement(2651..2670)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2652..2670)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2684..2700)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2684..2700)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2697..2719)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2708..2724
                     /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)."
     promoter        complement(join(2732..2738,2739..2756,2757..2762))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2777..2798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2915..2932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3086..3674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3166..3185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3845..4636,4637..4705))
                     /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     4468..4487
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4706..4810)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4837..5292
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4924..4943)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5134..5155
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5418..5440
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5432..5449
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5433..5449
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        join(5610..5774,1..215)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
       61 acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg
      121 gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc
      181 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca
      241 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      301 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg
      361 ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
      421 gtttagtgaa ccgtcagaat tgtttttatt tttaattttc tttcaaatac ttccatcgaa
      481 ttcagatctg gtaccacgcg taccatggat atcttttacc catacgatgt tcctgactat
      541 gcgggctatc cctatgacgt cccggactat gcaggatcct atccatatga cgttccagat
      601 tacgctgctc agatcgataa gcttatggcg gcgccggagg cggaggttct gtcctcagcc
      661 gcagtccctg atttggagtg gtatgagaag tccgaagaaa ctcacgcctc ccagatagaa
      721 ctacttgaga caagctctac gcaggaacct ctcaacgctt cggaggcctt ttgcccaaga
      781 gactgcatgg taccagtggt gtttcctggg cctgtgagcc aggaaggctg ctgtcagttt
      841 acttgtgaac ttctaaagca tatcatgtat caacgccagc agctccctct gccctatgaa
      901 cagcttaagc acttttaccg aaaaccttct ccccaggcag aggagatgct gaagaagaaa
      961 cctcgggcca ccactgaggt gagcagcagg aaatgccaac aagccctggc agaactggag
     1021 agtgtcctca gccacctgga ggacttcttt gcacggacac tagtaccgcg agtgctgatt
     1081 ctccttgggg gcaatgccct aagccccaag gagttctatg aactcgactt gtctctgctg
     1141 gccccctaca gcgtggacca gagcctgagc acagcagctt gtttgcgccg tctcttccga
     1201 gccatattca tggctgatgc ctttagcgag cttcaggctc ctccactcat gggcaccgtc
     1261 gtcatggcac agggacaccg caactgtgga gaagattggt ttcgacccaa gctcaactat
     1321 cgagtgccca gccggggcca taaactgact gtgaccctgt catgtggcag accttccatc
     1381 cgaaccacgg cttgggaaga ctacatttgg ttccaggcac cagtgacatt taaaggcttc
     1441 cgcgagtcta gaggatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     1501 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     1561 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     1621 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     1681 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     1741 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     1801 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     1861 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     1921 tcccttccct gtccttctga ttttaaaata actataccag caggaggacg tccagacaca
     1981 gcataggcta cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt
     2041 cctctcatgc gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcttg
     2101 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2161 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2221 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2281 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2341 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     2401 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcctcg aggaactgaa
     2461 aaaccagaaa gttaactggt aagtttagtc tttttgtctt ttatttcagg tcccggatcc
     2521 ggtggtggtg caaatcaaag aactgctcct cagtggatgt tgcctttact tctaggcctg
     2581 tacggaagtg ttacttctgc tctaaaagct gctgcaggtc gactctaggg ccgcgtaccg
     2641 agctcgaatt ccctatagtg agtcgtatta aattcgtaat catgtcatag ctgtttcctg
     2701 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     2761 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2821 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     2881 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2941 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3001 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3061 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3121 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3181 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3241 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3301 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3361 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3421 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3481 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3541 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3601 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3661 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3721 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3781 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3841 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3901 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3961 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4021 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4081 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4141 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4201 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4261 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4321 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4381 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4441 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4501 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4561 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4621 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4681 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4741 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4801 gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa
     4861 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4921 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4981 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5041 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5101 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5161 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     5221 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5281 cgcttacaat ttccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     5341 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     5401 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctgat
     5461 ctatacattg aatcaatatt ggcaattagc catattagtc attggttata tagcataaat
     5521 caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat
     5581 tggctcatgt ccaatatgac cgccatgttg acattgatta ttgactagtt attaatagta
     5641 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     5701 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     5761 gtatgttccc atag
//
