LOCUS       sequence-442594-        6578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Myc-tagged human RNF8 (R42A).
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-R42A
FEATURES             Location/Qualifiers
     source          1..6578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(11..79,80..871)
                     /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(229..248)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1042..1630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1531..1550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1784..1801
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1918..1939
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1954..1959,1960..1977,1978..1984)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1992..2008
                     /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     1997..2019
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2016..2032
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2016..2032
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2081..2202
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2092..2111)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2146..2165
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2332..2706)
                     /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"
     promoter        complement(2725..2772)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(2820..3149)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2834..2969
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2888..2907)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3134..3154
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3163..3591)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3273..3294)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3485..3504
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3637..3861
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3850..3867
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3890..3907)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3923..3952)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5560..5584)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    5594..5612
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5615..5633
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(5635..5838)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5664..5684)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5839..6218
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6390..6409
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(6484..6578,1..10)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
       61 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      121 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      181 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      241 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      301 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      361 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      421 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      481 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      541 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      601 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      661 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      721 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
      781 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
      841 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
      901 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
      961 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1021 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1081 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1141 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1201 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1261 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1321 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1381 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1441 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1501 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1561 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1621 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1681 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1741 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1801 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     1861 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     1921 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     1981 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     2041 cgccaagctc tagctagagg tcgacggtat acagacatga taagatacat tgatgagttt
     2101 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2161 attgctttat ttgtaaccat tataagctgc aataaacaag ttggggtggg cgaagaactc
     2221 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
     2281 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtagcacgt gtcagtcctg
     2341 ctcctcggcc acgaagtgca cgcagttgcc ggccgggtcg cgcagggcga actcccgccc
     2401 ccacggctgc tcgccgatct cggtcatggc cggcccggag gcgtcccgga agttcgtgga
     2461 cacgacctcc gaccactcgg cgtacagctc gtccaggccg cgcacccaca cccaggccag
     2521 ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg aacagggtca cgtcgtcccg
     2581 gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc ggtcggtcca
     2641 gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac tggtcaactt
     2701 ggccatggtt tagttcctca ccttgtcgta ttatactatg ccgatatact atgccgatga
     2761 ttaattgtca acacgtgctg atcagatccg aaaatggata tacaagctcc cgggagcttt
     2821 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggca
     2881 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
     2941 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
     3001 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     3061 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     3121 ggggactttc cacaccctaa ctgacacaca ttccacagaa ttaattcgcg ttaaattttt
     3181 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3241 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3301 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3361 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3421 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3481 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3541 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tggggatacc
     3601 ccctagagcc ccagctggtt ctttccgcct cagaagccat agagcccacc gcatccccag
     3661 catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca
     3721 gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga
     3781 cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg
     3841 gctggcaact agaaggcaca gtcgaggctg atcagcgggt ttaaactcaa tggtgatggt
     3901 gatgatgacc ggtatgcata ttcagatcct cttctgagat gagtttttgt tcgaagggcc
     3961 ctctagactc gaggaacaat ctctttgctt ttcgttccct aatgagaaca attcgtcgtt
     4021 ctttcacttc tgagctcaga ttatttacca tcttattaat gcaattgtcc agaaccaaag
     4081 agtatgtttt ggacttaatg tccttccgac aaatggggca ttctatcttc cgcttcatcc
     4141 attcattgat acagtaggag cagaaactgt gggcacagtt caaggtgaca gcctcaatga
     4201 agtattctga acaaataata cattggagct cattctctag cacatcattc atgtggctaa
     4261 gaacttcttc cttctgtgct tgcatcttct ccttctcttc cttggtctgc tctaattctt
     4321 tgttcttggc ttgaatgatt gcttcaaagt ccttcttgct gcgatttagc tcttccatta
     4381 gagcccaatg ctcctgcaga gcctgggcca gctgttgctt caggtccttt tctccttggg
     4441 ctatctccaa accctgaaga tgctgttcct cttcctggaa agtcttctct agttgctcca
     4501 ctcttgcctg ctgctgagcc tgctctgcac acagctggga ctgtaagtcc tgaagttcct
     4561 gctccatttg cacaactttc tttgagttcc ccttttgggt ctgctttttc acattcataa
     4621 cggcttcatg tttttcctgc atctgtattt tgagcctcag aatcctggac atggtcacct
     4681 taaacatctg taagcttctc tgagatgctg aagagtttga ggctttctgc tcatgatgaa
     4741 cctctgtgac tttggggaag ccagggtaaa tgggagcccc tgtggtctta cttggctcaa
     4801 gggcagtcaa cttaggatcc aaattgtcag agggtgtact ggccacttca cctttcccct
     4861 gtgatttcac tggctgacca gattcacaag acactttatt tattttggat ttcaaatttg
     4921 aggggccttc agctccagga cctgctaatt catccaaact gaatttcctt ttagttctca
     4981 attccttatt tttttctatc atttggtcat tctttgggga aagacaagga tatattgtct
     5041 cccagtcttc ttcagtaact tcatattcat actccgcatt ctccttattt tccagaggca
     5101 ctccaagttg gatgtagtct ccctgatgaa tggaatagac ccttaaaggt tccagacgcg
     5161 ctctgttcag ccaaacacca tttagactct tgttgtccat aattgtccat tggccctcag
     5221 gattctgctt caaaacacag tggtttcgag aaatcatcag ggggcagatt tttgatacca
     5281 gttggtatgt gacaccaaat cctgctccta cagtcacctc gcacccatct tccagcagca
     5341 gccacccggc gctcatcccc acccgccgca ggcaccagct ccggccaccg gcgcggtctc
     5401 ctgtgacgaa gaagccgggc tcccccatga attccaccac actggactag tggatccgag
     5461 ctcggtacca agcttaagtt taaacgctag agtccggagg ctggatcggt cccggtgtct
     5521 tctatggagg tcaaaacagc gtggatggcg tctccaggcg atctgacggt tcactaaacg
     5581 agctcgtcga cgatctctat cactgatagg gagatctcta tcactgatag ggagagctct
     5641 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     5701 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     5761 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     5821 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     5881 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     5941 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     6001 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     6061 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     6121 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     6181 aattgattac tattaataac tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac
     6241 atcgcgaagc agcgcaaaac gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa
     6301 gccttgcctt gttgtagctt aaattttgct cgcgcactac tcagcgacct ccaacacaca
     6361 agcagggagc agatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     6421 ccatagggga tcgggagatc tcccgatccg tcgacgtcag gtggcacttt tcggggaaat
     6481 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     6541 agacaataac cctgataaat gcttcaataa tattgaaa
//
