LOCUS       sequence-442605-        7838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Myc-tagged BirA*-RNF8 (C403S) fusion protein
            for BioID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7838)
  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 7838)
  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: pcDNA3.1-Myc-BirA*-RNF8-C403S
FEATURES             Location/Qualifiers
     source          1..7838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..40)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(194..782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(274..293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(953..1744,1745..1813))
                     /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     1576..1595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1814..1918)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1993..2012)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2184..2563
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2564..2767
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2718..2738
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2812..2831
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2812..2830
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2848..2857
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2857..2886
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(2890..3237,3238..3240,3241..3849)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     primer_bind     complement(5378..5395)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5384..5608
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5654..6082
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5741..5760)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5951..5972
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6091..6111)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6096..6425
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6276..6411
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6338..6357
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6492..7286
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6546..6565)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7156..7175
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7460..7581
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7497..7516)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7551..7570
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7630..7646)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7630..7646)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7643..7665)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7654..7670
                     /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(7678..7684,7685..7702,7703..7708))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7723..7744
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
       61 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      121 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      181 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      241 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      301 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      361 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      421 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      481 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      541 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      601 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
      661 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      721 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      781 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      841 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      901 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
      961 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1021 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1081 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1141 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1201 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1261 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1321 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1381 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1441 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1501 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1561 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1621 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1681 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1741 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1801 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1861 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1921 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     1981 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     2041 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     2101 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     2161 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
     2221 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     2281 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     2341 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     2401 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     2461 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     2521 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     2581 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     2641 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     2701 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     2761 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
     2821 tcactatagg gagacccaag ctggctagcc accatggaac aaaaactcat ctcagaagag
     2881 gatctcgaca aggacaacac cgtgcccctg aagctgatcg ccctgctggc caacggcgag
     2941 ttccactctg gcgagcagct gggagagacc ctgggaatga gcagagccgc catcaacaag
     3001 cacatccaga cactgagaga ctggggagtg gacgtgttca ccgtgcctgg caagggctac
     3061 agcctgcctg agcctatcca gctgctgaac gccaagcaga tcctgggaca gctggatggc
     3121 ggaagcgtgg ccgtgctgcc tgtgatcgac tccaccaatc agtacctgct ggacagaatc
     3181 ggagagctga agtccggcga cgcctgcatc gccgagtacc agcaggctgg cagaggaggc
     3241 agaggacgga agtggttcag cccattcgga gccaacctgt acctgtccat gttctggaga
     3301 ctggagcagg gacctgctgc tgccatcgga ctgagtctgg tgatcggaat cgtgatggcc
     3361 gaggtgctga gaaagctggg agccgacaag gtgagagtga agtggcctaa tgacctgtac
     3421 ctccaggacc gcaagctggc tggcatcctg gtggagctga caggcaagac aggcgatgcc
     3481 gctcagatcg tgatcggagc cggaatcaac atggccatga gaagagtgga ggagagcgtg
     3541 gtgaaccagg gctggatcac cctgcaggag gctggcatca acctggaccg gaacaccctg
     3601 gccgccatgc tgatcagaga gctgagagcc gctctggagc tgttcgagca ggagggactg
     3661 gctccttacc tgagcagatg ggagaagctg gacaacttca tcaacagacc tgtgaagctg
     3721 atcatcggcg acaaggaaat cttcggcatc tccagaggaa tcgacaagca gggagctctg
     3781 ctgctggagc aggacggaat catcaagccc tggatgggcg gagaaatctc cctgagaagc
     3841 gcagagaagc tcgaggggga gcccggcttc ttcgtcacag gagaccgcgc cggtggccgg
     3901 agctggtgcc tgcggcgggt ggggatgagc gccgggtggc tgctgctgga agatgggtgc
     3961 gaggtgactg taggacgagg atttggtgtc acataccaac tggtatcaaa aatctgcccc
     4021 ctgatgattt ctcgaaacca ctgtgttttg aagcagaatc ctgagggcca atggacaatt
     4081 atggacaaca agagtctaaa tggtgtttgg ctgaacagag cgcgtctgga acctttaagg
     4141 gtctattcca ttcatcaggg agactacatc caacttggag tgcctctgga aaataaggag
     4201 aatgcggagt atgaatatga agttactgaa gaagactggg agacaatata tccttgtctt
     4261 tccccaaaga atgaccaaat gatagaaaaa aataaggaat tgagaactaa aaggaaattc
     4321 agtttggatg aattagcagg tcctggagct gaaggcccct caaatttgaa atccaaaata
     4381 aataaagtgt cttgtgaatc tggtcagcca gtgaaatcac aggggaaagg tgaagtggcc
     4441 agtacaccct ctgacaattt ggatcctaag ttgactgccc ttgagccaag taagaccaca
     4501 ggggctccca tttaccctgg cttccccaaa gtcacagagg ttcatcatga gcagaaagcc
     4561 tcaaactctt cagcatctca gagaagctta cagatgttta aggtgaccat gtccaggatt
     4621 ctgaggctca aaatacagat gcaggaaaaa catgaagccg ttatgaatgt gaaaaagcag
     4681 acccaaaagg ggaactcaaa gaaagttgtg caaatggagc aggaacttca ggacttacag
     4741 tcccagctgt gtgcagagca ggctcagcag caggcaagag tggagcaact agagaagact
     4801 ttccaggaag aggaacagca tcttcagggt ttggagatag cccaaggaga aaaggacctg
     4861 aagcaacagc tggcccaggc tctgcaggag cattgggctc taatggaaga gctaaatcgc
     4921 agcaagaagg actttgaagc aatcattcaa gccaagaaca aagaattaga gcagaccaag
     4981 gaagagaagg agaagatgca agcacagaag gaagaagttc ttagccacat gaatgatgtg
     5041 ctagagaatg agctccaaag tattatttgt tcagaatact tcattgaggc tgtcaccttg
     5101 aactgtgccc acagtttctg ctcctactgt atcaatgaat ggatgaagcg gaagatagaa
     5161 tgccccattt gtcggaagga cattaagtcc aaaacatact ctttggttct ggacaattgc
     5221 attaataaga tggtaaataa tctgagctca gaagtgaaag aacgacgaat tgttctcatt
     5281 agggaacgaa aagcaaagag attgttctga gaattccacc acactggact agtggatccg
     5341 agctcggtac caagcttaag tttaaaccgc tgatcagcct cgactgtgcc ttctagttgc
     5401 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     5461 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     5521 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     5581 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     5641 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5701 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5761 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5821 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     5881 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5941 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     6001 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     6061 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     6121 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     6181 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     6241 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     6301 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     6361 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     6421 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     6481 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     6541 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     6601 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     6661 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     6721 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     6781 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     6841 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     6901 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     6961 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     7021 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     7081 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     7141 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     7201 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     7261 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     7321 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     7381 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     7441 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     7501 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     7561 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     7621 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     7681 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     7741 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     7801 ttaatgaatc ggccaacgcg cggggagagg cggtttgc
//
