LOCUS       sequence-442598-        6578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Myc-tagged human RNF8 (T198A).
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-T198A
FEATURES             Location/Qualifiers
     source          1..6578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      647..1235
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1136..1155
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1389..1406
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1523..1544
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1559..1564,1565..1582,1583..1589)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1597..1613
                     /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     1602..1624
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1621..1637
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1621..1637
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1686..1807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1697..1716)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1751..1770
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1937..2311)
                     /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(2330..2377)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(2425..2754)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2439..2574
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2493..2512)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2739..2759
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2768..3196)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2878..2899)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3090..3109
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3242..3466
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3455..3472
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3495..3512)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3528..3557)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5165..5189)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    5199..5217
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5220..5238
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(5240..5443)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5269..5289)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5444..5823
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5995..6014
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6089..6193
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6194..6262,6263..6578,1..476)
                     /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(6412..6431)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
       61 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
      121 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
      181 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
      241 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
      301 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
      361 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
      421 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
      481 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
      541 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
      601 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
      661 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
      721 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
      781 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
      841 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
      901 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
      961 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1021 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1081 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1141 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     1201 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     1261 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     1321 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     1381 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     1441 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     1501 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     1561 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     1621 caggaaacag ctatgaccat gattacgcca agctctagct agaggtcgac ggtatacaga
     1681 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     1741 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     1801 acaagttggg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
     1861 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
     1921 aatctcgtag cacgtgtcag tcctgctcct cggccacgaa gtgcacgcag ttgccggccg
     1981 ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc gatctcggtc atggccggcc
     2041 cggaggcgtc ccggaagttc gtggacacga cctccgacca ctcggcgtac agctcgtcca
     2101 ggccgcgcac ccacacccag gccagggtgt tgtccggcac cacctggtcc tggaccgcgc
     2161 tgatgaacag ggtcacgtcg tcccggacca caccggcgaa gtcgtcctcc acgaagtccc
     2221 gggagaaccc gagccggtcg gtccagaact cgaccgctcc ggcgacgtcg cgcgcggtga
     2281 gcaccggaac ggcactggtc aacttggcca tggtttagtt cctcaccttg tcgtattata
     2341 ctatgccgat atactatgcc gatgattaat tgtcaacacg tgctgatcag atccgaaaat
     2401 ggatatacaa gctcccggga gctttttgca aaagcctagg cctccaaaaa agcctcctca
     2461 ctacttctgg aatagctcag aggcagaggc ggcctcggcc tctgcataaa taaaaaaaat
     2521 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     2581 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     2641 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
     2701 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
     2761 cagaattaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     2821 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     2881 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     2941 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3001 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3061 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3121 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3181 ccgctacagg gcgcgtgggg atacccccta gagccccagc tggttctttc cgcctcagaa
     3241 gccatagagc ccaccgcatc cccagcatgc ctgctattgt cttcccaatc ctcccccttg
     3301 ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca
     3361 atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc
     3421 acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga ggctgatcag
     3481 cgggtttaaa ctcaatggtg atggtgatga tgaccggtat gcatattcag atcctcttct
     3541 gagatgagtt tttgttcgaa gggccctcta gactcgagga acaatctctt tgcttttcgt
     3601 tccctaatga gaacaattcg tcgttctttc acttctgagc tcagattatt taccatctta
     3661 ttaatgcaat tgtccagaac caaagagtat gttttggact taatgtcctt ccgacaaatg
     3721 gggcattcta tcttccgctt catccattca ttgatacagt aggagcagaa actgtgggca
     3781 cagttcaagg tgacagcctc aatgaagtat tctgaacaaa taatacattg gagctcattc
     3841 tctagcacat cattcatgtg gctaagaact tcttccttct gtgcttgcat cttctccttc
     3901 tcttccttgg tctgctctaa ttctttgttc ttggcttgaa tgattgcttc aaagtccttc
     3961 ttgctgcgat ttagctcttc cattagagcc caatgctcct gcagagcctg ggccagctgt
     4021 tgcttcaggt ccttttctcc ttgggctatc tccaaaccct gaagatgctg ttcctcttcc
     4081 tggaaagtct tctctagttg ctccactctt gcctgctgct gagcctgctc tgcacacagc
     4141 tgggactgta agtcctgaag ttcctgctcc atttgcacaa ctttctttga gttccccttt
     4201 tgggtctgct ttttcacatt cataacggct tcatgttttt cctgcatctg tattttgagc
     4261 ctcagaatcc tggacatggt caccttaaac atctgtaagc ttctctgaga tgctgaagag
     4321 tttgaggctt tctgctcatg atgaacctct gtgactttgg ggaagccagg gtaaatggga
     4381 gcccctgtgg tcttacttgg ctcaagggca gtcaacttag gatccaaatt gtcagagggt
     4441 gcactggcca cttcaccttt cccctgtgat ttcactggct gaccagattc acaagacact
     4501 ttatttattt tggatttcaa atttgagggg ccttcagctc caggacctgc taattcatcc
     4561 aaactgaatt tccttttagt tctcaattcc ttattttttt ctatcatttg gtcattcttt
     4621 ggggaaagac aaggatatat tgtctcccag tcttcttcag taacttcata ttcatactcc
     4681 gcattctcct tattttccag aggcactcca agttggatgt agtctccctg atgaatggaa
     4741 tagaccctta aaggttccag acgcgctctg ttcagccaaa caccatttag actcttgttg
     4801 tccataattg tccattggcc ctcaggattc tgcttcaaaa cacagtggtt tcgagaaatc
     4861 atcagggggc agatttttga taccagttgg tatgtgacac caaatcctcg tcctacagtc
     4921 acctcgcacc catcttccag cagcagccac ccggcgctca tccccacccg ccgcaggcac
     4981 cagctccggc caccggcgcg gtctcctgtg acgaagaagc cgggctcccc catgaattcc
     5041 accacactgg actagtggat ccgagctcgg taccaagctt aagtttaaac gctagagtcc
     5101 ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa acagcgtgga tggcgtctcc
     5161 aggcgatctg acggttcact aaacgagctc gtcgacgatc tctatcactg atagggagat
     5221 ctctatcact gatagggaga gctctgctta tatagacctc ccaccgtaca cgcctaccgc
     5281 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt
     5341 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa
     5401 accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga
     5461 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     5521 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     5581 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     5641 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     5701 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     5761 atgggctatg aactaatgac cccgtaattg attactatta ataactagtc aataatcaat
     5821 gtcaacgcgt atatctggcc cgtacatcgc gaagcagcgc aaaacgccta accctaagca
     5881 gattcttcat gcaattgtcg gtcaagcctt gccttgttgt agcttaaatt ttgctcgcgc
     5941 actactcagc gacctccaac acacaagcag ggagcagata ctggcttaac tatgcggcat
     6001 cagagcagat tgtactgaga gtgcaccata ggggatcggg agatctcccg atccgtcgac
     6061 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6121 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6181 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6241 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6301 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6361 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     6421 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6481 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     6541 agtaagagaa ttatgcagtg ctgccataac catgagtg
//
