LOCUS       sequence-440443-        7838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Myc-tagged BirA*-RNF8 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
FEATURES             Location/Qualifiers
     source          1..7838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(11..599)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(91..110)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(770..1561,1562..1630))
                     /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     1393..1412
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1631..1735)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1810..1829)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2001..2380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2381..2584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2535..2555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2629..2648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2629..2647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2665..2674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2674..2703
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(2707..3054,3055..3057,3058..3666)
                     /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(5195..5212)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5201..5425
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5471..5899
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5558..5577)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5768..5789
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5908..5928)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5913..6242
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6093..6228
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6155..6174
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6309..7103
                     /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(6363..6382)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6973..6992
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7277..7398
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7314..7333)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7368..7387
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7447..7463)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7447..7463)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7460..7482)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7471..7487
                     /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(7495..7501,7502..7519,7520..7525))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7540..7561
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7678..7695)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
       61 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
      121 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      181 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
      241 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
      301 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
      361 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
      421 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
      481 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
      541 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
      601 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
      661 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
      721 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
      781 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
      841 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
      901 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
      961 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     1021 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1081 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1141 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1201 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     1261 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     1321 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     1381 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     1441 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     1501 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     1561 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     1621 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     1681 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     1741 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
     1801 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     1861 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     1921 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
     1981 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
     2041 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     2101 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     2161 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     2221 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     2281 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     2341 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     2401 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     2461 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     2521 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     2581 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
     2641 ctatagggag acccaagctg gctagccacc atggaacaaa aactcatctc agaagaggat
     2701 ctcgacaagg acaacaccgt gcccctgaag ctgatcgccc tgctggccaa cggcgagttc
     2761 cactctggcg agcagctggg agagaccctg ggaatgagca gagccgccat caacaagcac
     2821 atccagacac tgagagactg gggagtggac gtgttcaccg tgcctggcaa gggctacagc
     2881 ctgcctgagc ctatccagct gctgaacgcc aagcagatcc tgggacagct ggatggcgga
     2941 agcgtggccg tgctgcctgt gatcgactcc accaatcagt acctgctgga cagaatcgga
     3001 gagctgaagt ccggcgacgc ctgcatcgcc gagtaccagc aggctggcag aggaggcaga
     3061 ggacggaagt ggttcagccc attcggagcc aacctgtacc tgtccatgtt ctggagactg
     3121 gagcagggac ctgctgctgc catcggactg agtctggtga tcggaatcgt gatggccgag
     3181 gtgctgagaa agctgggagc cgacaaggtg agagtgaagt ggcctaatga cctgtacctc
     3241 caggaccgca agctggctgg catcctggtg gagctgacag gcaagacagg cgatgccgct
     3301 cagatcgtga tcggagccgg aatcaacatg gccatgagaa gagtggagga gagcgtggtg
     3361 aaccagggct ggatcaccct gcaggaggct ggcatcaacc tggaccggaa caccctggcc
     3421 gccatgctga tcagagagct gagagccgct ctggagctgt tcgagcagga gggactggct
     3481 ccttacctga gcagatggga gaagctggac aacttcatca acagacctgt gaagctgatc
     3541 atcggcgaca aggaaatctt cggcatctcc agaggaatcg acaagcaggg agctctgctg
     3601 ctggagcagg acggaatcat caagccctgg atgggcggag aaatctccct gagaagcgca
     3661 gagaagctcg agggggagcc cggcttcttc gtcacaggag accgcgccgg tggccggagc
     3721 tggtgcctgc ggcgggtggg gatgagcgcc gggtggctgc tgctggaaga tgggtgcgag
     3781 gtgactgtag gacgaggatt tggtgtcaca taccaactgg tatcaaaaat ctgccccctg
     3841 atgatttctc gaaaccactg tgttttgaag cagaatcctg agggccaatg gacaattatg
     3901 gacaacaaga gtctaaatgg tgtttggctg aacagagcgc gtctggaacc tttaagggtc
     3961 tattccattc atcagggaga ctacatccaa cttggagtgc ctctggaaaa taaggagaat
     4021 gcggagtatg aatatgaagt tactgaagaa gactgggaga caatatatcc ttgtctttcc
     4081 ccaaagaatg accaaatgat agaaaaaaat aaggaattga gaactaaaag gaaattcagt
     4141 ttggatgaat tagcaggtcc tggagctgaa ggcccctcaa atttgaaatc caaaataaat
     4201 aaagtgtctt gtgaatctgg tcagccagtg aaatcacagg ggaaaggtga agtggccagt
     4261 acaccctctg acaatttgga tcctaagttg actgcccttg agccaagtaa gaccacaggg
     4321 gctcccattt accctggctt ccccaaagtc acagaggttc atcatgagca gaaagcctca
     4381 aactcttcag catctcagag aagcttacag atgtttaagg tgaccatgtc caggattctg
     4441 aggctcaaaa tacagatgca ggaaaaacat gaagccgtta tgaatgtgaa aaagcagacc
     4501 caaaagggga actcaaagaa agttgtgcaa atggagcagg aacttcagga cttacagtcc
     4561 cagctgtgtg cagagcaggc tcagcagcag gcaagagtgg agcaactaga gaagactttc
     4621 caggaagagg aacagcatct tcagggtttg gagatagccc aaggagaaaa ggacctgaag
     4681 caacagctgg cccaggctct gcaggagcat tgggctctaa tggaagagct aaatcgcagc
     4741 aagaaggact ttgaagcaat cattcaagcc aagaacaaag aattagagca gaccaaggaa
     4801 gagaaggaga agatgcaagc acagaaggaa gaagttctta gccacatgaa tgatgtgcta
     4861 gagaatgagc tccaatgtat tatttgttca gaatacttca ttgaggctgt caccttgaac
     4921 tgtgcccaca gtttctgctc ctactgtatc aatgaatgga tgaagcggaa gatagaatgc
     4981 cccatttgtc ggaaggacat taagtccaaa acatactctt tggttctgga caattgcatt
     5041 aataagatgg taaataatct gagctcagaa gtgaaagaac gacgaattgt tctcattagg
     5101 gaacgaaaag caaagagatt gttctgagaa ttccaccaca ctggactagt ggatccgagc
     5161 tcggtaccaa gcttaagttt aaaccgctga tcagcctcga ctgtgccttc tagttgccag
     5221 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     5281 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     5341 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     5401 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     5461 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     5521 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     5581 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     5641 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     5701 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5761 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     5821 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     5881 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     5941 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     6001 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     6061 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     6121 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     6181 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     6241 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     6301 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     6361 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     6421 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     6481 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     6541 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     6601 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     6661 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     6721 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     6781 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     6841 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     6901 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     6961 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     7021 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     7081 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     7141 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     7201 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     7261 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     7321 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     7381 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     7441 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7501 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7561 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7621 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7681 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7741 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7801 aggccagcaa aaggccagga accgtaaaaa ggccgcgt
//
