LOCUS       sequence-442591-        6578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Myc-tagged human RNF8.
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
FEATURES             Location/Qualifiers
     source          1..6578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(133..507)
                     /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(526..573)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(621..950)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      635..770
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(689..708)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     935..955
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(964..1392)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1074..1095)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1286..1305
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    1438..1662
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1651..1668
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1691..1708)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1724..1753)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3361..3385)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3395..3413
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3416..3434
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(3436..3639)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3465..3485)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3640..4019
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4191..4210
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4285..4389
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4390..4458,4459..5250)
                     /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(4608..4627)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5421..6009
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5910..5929
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6163..6180
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6297..6318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6333..6338,6339..6356,6357..6363)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6371..6387
                     /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     6376..6398
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6395..6411
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6395..6411
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    join(6460..6578,1..3)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6471..6490)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6525..6544
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gttggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc
       61 gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc
      121 tcgtagcacg tgtcagtcct gctcctcggc cacgaagtgc acgcagttgc cggccgggtc
      181 gcgcagggcg aactcccgcc cccacggctg ctcgccgatc tcggtcatgg ccggcccgga
      241 ggcgtcccgg aagttcgtgg acacgacctc cgaccactcg gcgtacagct cgtccaggcc
      301 gcgcacccac acccaggcca gggtgttgtc cggcaccacc tggtcctgga ccgcgctgat
      361 gaacagggtc acgtcgtccc ggaccacacc ggcgaagtcg tcctccacga agtcccggga
      421 gaacccgagc cggtcggtcc agaactcgac cgctccggcg acgtcgcgcg cggtgagcac
      481 cggaacggca ctggtcaact tggccatggt ttagttcctc accttgtcgt attatactat
      541 gccgatatac tatgccgatg attaattgtc aacacgtgct gatcagatcc gaaaatggat
      601 atacaagctc ccgggagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
      661 ttctggaata gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt
      721 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
      781 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
      841 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata
      901 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacaga
      961 attaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1021 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1081 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1141 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     1201 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     1261 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     1321 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     1381 tacagggcgc gtggggatac cccctagagc cccagctggt tctttccgcc tcagaagcca
     1441 tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt
     1501 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt
     1561 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
     1621 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct gatcagcggg
     1681 tttaaactca atggtgatgg tgatgatgac cggtatgcat attcagatcc tcttctgaga
     1741 tgagtttttg ttcgaagggc cctctagact cgaggaacaa tctctttgct tttcgttccc
     1801 taatgagaac aattcgtcgt tctttcactt ctgagctcag attatttacc atcttattaa
     1861 tgcaattgtc cagaaccaaa gagtatgttt tggacttaat gtccttccga caaatggggc
     1921 attctatctt ccgcttcatc cattcattga tacagtagga gcagaaactg tgggcacagt
     1981 tcaaggtgac agcctcaatg aagtattctg aacaaataat acattggagc tcattctcta
     2041 gcacatcatt catgtggcta agaacttctt ccttctgtgc ttgcatcttc tccttctctt
     2101 ccttggtctg ctctaattct ttgttcttgg cttgaatgat tgcttcaaag tccttcttgc
     2161 tgcgatttag ctcttccatt agagcccaat gctcctgcag agcctgggcc agctgttgct
     2221 tcaggtcctt ttctccttgg gctatctcca aaccctgaag atgctgttcc tcttcctgga
     2281 aagtcttctc tagttgctcc actcttgcct gctgctgagc ctgctctgca cacagctggg
     2341 actgtaagtc ctgaagttcc tgctccattt gcacaacttt ctttgagttc cccttttggg
     2401 tctgcttttt cacattcata acggcttcat gtttttcctg catctgtatt ttgagcctca
     2461 gaatcctgga catggtcacc ttaaacatct gtaagcttct ctgagatgct gaagagtttg
     2521 aggctttctg ctcatgatga acctctgtga ctttggggaa gccagggtaa atgggagccc
     2581 ctgtggtctt acttggctca agggcagtca acttaggatc caaattgtca gagggtgtac
     2641 tggccacttc acctttcccc tgtgatttca ctggctgacc agattcacaa gacactttat
     2701 ttattttgga tttcaaattt gaggggcctt cagctccagg acctgctaat tcatccaaac
     2761 tgaatttcct tttagttctc aattccttat ttttttctat catttggtca ttctttgggg
     2821 aaagacaagg atatattgtc tcccagtctt cttcagtaac ttcatattca tactccgcat
     2881 tctccttatt ttccagaggc actccaagtt ggatgtagtc tccctgatga atggaataga
     2941 cccttaaagg ttccagacgc gctctgttca gccaaacacc atttagactc ttgttgtcca
     3001 taattgtcca ttggccctca ggattctgct tcaaaacaca gtggtttcga gaaatcatca
     3061 gggggcagat ttttgatacc agttggtatg tgacaccaaa tcctcgtcct acagtcacct
     3121 cgcacccatc ttccagcagc agccacccgg cgctcatccc cacccgccgc aggcaccagc
     3181 tccggccacc ggcgcggtct cctgtgacga agaagccggg ctcccccatg aattccacca
     3241 cactggacta gtggatccga gctcggtacc aagcttaagt ttaaacgcta gagtccggag
     3301 gctggatcgg tcccggtgtc ttctatggag gtcaaaacag cgtggatggc gtctccaggc
     3361 gatctgacgg ttcactaaac gagctcgtcg acgatctcta tcactgatag ggagatctct
     3421 atcactgata gggagagctc tgcttatata gacctcccac cgtacacgcc taccgcccat
     3481 ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca
     3541 aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg
     3601 ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa
     3661 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca
     3721 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
     3781 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
     3841 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
     3901 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg
     3961 gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtca
     4021 acgcgtatat ctggcccgta catcgcgaag cagcgcaaaa cgcctaaccc taagcagatt
     4081 cttcatgcaa ttgtcggtca agccttgcct tgttgtagct taaattttgc tcgcgcacta
     4141 ctcagcgacc tccaacacac aagcagggag cagatactgg cttaactatg cggcatcaga
     4201 gcagattgta ctgagagtgc accatagggg atcgggagat ctcccgatcc gtcgacgtca
     4261 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4321 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4381 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4441 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4501 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4561 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4621 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4681 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4741 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4801 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4861 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4921 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4981 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5041 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5101 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5161 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5221 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5281 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5341 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5401 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5461 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5521 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5581 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5641 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5701 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5761 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5821 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5881 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5941 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6001 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6061 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6121 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6181 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6241 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6301 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     6361 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     6421 gccaagctct agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg
     6481 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     6541 ttgctttatt tgtaaccatt ataagctgca ataaacaa
//
