LOCUS       sequence-442599-        6578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Myc-tagged human RNF8 (R477A/E478A/R479A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6578)
  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 6578)
  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: pcDNA4.1-Myc-RNF8-3A
FEATURES             Location/Qualifiers
     source          1..6578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(115..219)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(294..313)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        485..864
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        865..1068
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1019..1039
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    1070..1088
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1091..1109
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1119..1143
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2751..2780
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2796..2813
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2836..2853)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2842..3066
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3112..3540
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3199..3218)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3409..3430
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3549..3569)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3554..3883
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3734..3869
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3796..3815
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3931..3978
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3997..4371
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4501..4622
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4538..4557)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4592..4611
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4671..4687)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4671..4687)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4684..4706)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4695..4711
                     /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(4719..4725,4726..4743,4744..4749))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4764..4785
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4902..4919)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5073..5661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5153..5172)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5832..6578,1..45,46..114))
                     /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     6455..6474
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
       61 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
      121 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
      181 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
      241 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
      301 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
      361 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
      421 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
      481 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
      541 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      601 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      661 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      721 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      781 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      841 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      901 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      961 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     1021 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt
     1081 gatagagatc tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag
     1141 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
     1201 agcctccgga ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca
     1261 gtgtggtgga attcatgggg gagcccggct tcttcgtcac aggagaccgc gccggtggcc
     1321 ggagctggtg cctgcggcgg gtggggatga gcgccgggtg gctgctgctg gaagatgggt
     1381 gcgaggtgac tgtaggacga ggatttggtg tcacatacca actggtatca aaaatctgcc
     1441 ccctgatgat ttctcgaaac cactgtgttt tgaagcagaa tcctgagggc caatggacaa
     1501 ttatggacaa caagagtcta aatggtgttt ggctgaacag agcgcgtctg gaacctttaa
     1561 gggtctattc cattcatcag ggagactaca tccaacttgg agtgcctctg gaaaataagg
     1621 agaatgcgga gtatgaatat gaagttactg aagaagactg ggagacaata tatccttgtc
     1681 tttccccaaa gaatgaccaa atgatagaaa aaaataagga attgagaact aaaaggaaat
     1741 tcagtttgga tgaattagca ggtcctggag ctgaaggccc ctcaaatttg aaatccaaaa
     1801 taaataaagt gtcttgtgaa tctggtcagc cagtgaaatc acaggggaaa ggtgaagtgg
     1861 ccagtacacc ctctgacaat ttggatccta agttgactgc ccttgagcca agtaagacca
     1921 caggggctcc catttaccct ggcttcccca aagtcacaga ggttcatcat gagcagaaag
     1981 cctcaaactc ttcagcatct cagagaagct tacagatgtt taaggtgacc atgtccagga
     2041 ttctgaggct caaaatacag atgcaggaaa aacatgaagc cgttatgaat gtgaaaaagc
     2101 agacccaaaa ggggaactca aagaaagttg tgcaaatgga gcaggaactt caggacttac
     2161 agtcccagct gtgtgcagag caggctcagc agcaggcaag agtggagcaa ctagagaaga
     2221 ctttccagga agaggaacag catcttcagg gtttggagat agcccaagga gaaaaggacc
     2281 tgaagcaaca gctggcccag gctctgcagg agcattgggc tctaatggaa gagctaaatc
     2341 gcagcaagaa ggactttgaa gcaatcattc aagccaagaa caaagaatta gagcagacca
     2401 aggaagagaa ggagaagatg caagcacaga aggaagaagt tcttagccac atgaatgatg
     2461 tgctagagaa tgagctccaa tgtattattt gttcagaata cttcattgag gctgtcacct
     2521 tgaactgtgc ccacagtttc tgctcctact gtatcaatga atggatgaag cggaagatag
     2581 aatgccccat ttgtcggaag gacattaagt ccaaaacata ctctttggtt ctggacaatt
     2641 gcattaataa gatggtaaat aatctgagct cagaagtgaa agaacgacga attgttctca
     2701 ttgcggctgc aaaagcaaag agattgttcc tcgagtctag agggcccttc gaacaaaaac
     2761 tcatctcaga agaggatctg aatatgcata ccggtcatca tcaccatcac cattgagttt
     2821 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2881 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2941 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3001 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     3061 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     3121 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3181 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3241 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3301 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3361 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3421 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3481 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3541 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3601 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3661 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3721 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3781 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     3841 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     3901 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
     3961 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt
     4021 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
     4081 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
     4141 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
     4201 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
     4261 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
     4321 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta
     4381 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4441 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4501 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4561 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4621 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4681 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4741 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4801 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4861 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4921 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4981 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5041 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5101 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5161 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5221 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5281 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5341 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5401 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5461 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5521 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5581 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5641 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5701 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5761 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5821 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5881 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5941 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6001 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6061 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6121 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6181 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6241 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6301 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6361 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6421 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6481 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6541 ctgttgagat ccagttcgat gtaacccact cgtgcacc
//
