LOCUS       sequence-442607-        5570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of HA-tagged human MAD2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5570)
  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 5570)
  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-MAD2
FEATURES             Location/Qualifiers
     source          1..5570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     255..275
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    299..921
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     703..722
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(1571..1660)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     promoter        complement(1758..1961)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1787..1807)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1962..2341
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(2502..2519)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2502..2518)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2511..2533)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2659..3114)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2796..2817)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3008..3027
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3141..3245
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3246..3314,3315..4106)
                     /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(3464..3483)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4277..4865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4766..4785
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5019..5036
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5153..5174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5189..5194,5195..5212,5213..5219)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5227..5243
                     /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     5232..5254
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5251..5267
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5251..5267
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5281..5300
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5281..5299
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          5376..5472
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     promoter        complement(join(5511..5570,1..270))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5525..5570,1..90)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
       61 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
      121 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
      181 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
      241 tggggacttt ccacacccta actgacacac attccacagc caagctggcc gcgtacccaa
      301 ttcaacaggc atctactgag tggacccaac gcatgagagg acagtgccaa gcaagcaact
      361 caaatgtccc accggttggg catggccagg tagcctatgc tgtgtctgga cgtcctcctg
      421 ctggtatagt tattttaaaa tcagaaggac agggaaggga gcagtggttc acgcctgtaa
      481 tcccagcaat ttgggaggcc aaggtgggta gatcacctga gattaggagt tggagaccag
      541 cctggccaat atggtgaaac cccgtctcta ccaaaaaaac aaaaattagc tgagcctggt
      601 catgcatgcc tggaatccca acaactcggg aggctgaggc aggagaatcg cttgaaccca
      661 ggaggcggag attgcagtga gccaagattg tgccactgca ctccagcttg gttcccaata
      721 gaccccgcag gccctacagg ttgtcttccc aacttgcccc ttgctccata ccacccccct
      781 ccaccccata atattataga aggacaccta gtcagacaaa atgatgcaac ttaattttat
      841 taggacaagg ctggtgggca ctggagtggc aacttccagg gccaggagag gcactgggga
      901 ggggtcacag ggatgccacc cgggatcctc tagatcagtc attgacagga attttgtagg
      961 ccaccatgct atttactttg tggattgtag tagtaaatga acgaaggcgg acttcctcag
     1021 aattggtaat aaactgtggt cccgactctt cccatttttc aggtacaacc aaatctttgt
     1081 ctgtataaat cagcagatca aatgaacaag aaacttccaa cagtggcaga aatgtcaccg
     1141 tagctgtgat ctgtctgatc actgaacgga tttcatcctg gatagctttc tgagactttt
     1201 ctctgggtgc actgtcatct tttgcagtct tgtcacactc aatatcaaac tgccatcttt
     1261 ccaggacctc accactttca atatttgaga taactacaac cagtttctga actgaacact
     1321 tgtataacca atctttcagt tgttccacca cattatttag gtattttatg agctcaagat
     1381 cagtagttac aagcaaggtg agtccgtatt tctgcactcg agtaaaggtt tcagatggat
     1441 atatgccacg ctgatataaa atgctgttga tgccgaatga gaagaactcg gccacgattt
     1501 cggcgctccc gcgcagggtg attccctgct cccgggagag ctgcagcgcc ataagcttat
     1561 cgatctgagc agcgtaatct ggaacgtcat atggatagga tcctgcatag tccgggacgt
     1621 catagggata gcccgcatag tcaggaacat cgtatgggta aaagatatcc atggtacgcg
     1681 tggtaccaga tctgaattcg atggaagtat ttgaaagaaa attaaaaata aaaacaattc
     1741 tgacggttca ctaaacgagc tctgcttata tagacctccc accgtacacg cctaccgccc
     1801 atttgcgtca acggggcggg gttattacga cattttggaa agtcccgttg attttggtgc
     1861 caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac
     1921 cgctatccac gcccattggt gtactgccaa aaccgcatca ccatggtaat agcgatgact
     1981 aatacgtaga tgtactgcca agtaggaaag tcccgtaagg tcatgtactg ggcataatgc
     2041 caggcgggcc atttaccgtc attgacgtca atagggggcg gacttggcat atgatacact
     2101 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
     2161 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
     2221 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
     2281 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt
     2341 caacatggcg gtcatattgg acatgagcca atataaatgt acatattatg atatagatac
     2401 aacgtatgca atggccaata gccaatattg atttatgcta tataaccaat gactaatatg
     2461 gctaattgcc aatattgatt caatgtatag atcagcttgg cactggccgt cgttttacaa
     2521 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     2581 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     2641 agcctgaatg gcgaatggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     2701 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2761 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     2821 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     2881 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2941 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3001 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3061 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3121 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3181 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3241 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3301 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3361 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3421 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3481 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3541 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3601 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3661 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3721 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3781 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3841 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3901 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3961 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4021 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4081 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4141 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4201 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4261 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4321 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4381 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4441 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4501 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4561 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4621 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4681 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4741 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4801 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4861 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4921 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4981 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5041 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     5101 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     5161 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     5221 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgacatg attacgaatt
     5281 taatacgact cactataggg aattcgagct cggtacgcgg ccctagagtc gacctgcagc
     5341 agcttttaga gcagaagtaa cacttccgta caggcctaga agtaaaggca acatccactg
     5401 aggagcagtt ctttgatttg caccaccacc ggatccggga cctgaaataa aagacaaaaa
     5461 gactaaactt accagttaac tttctggttt ttcagttcct cgaggagctt tttgcaaaag
     5521 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
//
