LOCUS       sequence-446877-        7897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Myc-tagged BirA*-CAMK2D fusion protein for 
            BioID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7897)
  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 7897)
  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*-CAMK2D
FEATURES             Location/Qualifiers
     source          1..7897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(938..967)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      967..976
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(993..1012)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(994..1012)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1057..1260)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1086..1106)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1261..1640
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1812..1831
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1906..2010
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2011..2079,2080..2871)
                     /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(2229..2248)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3042..3630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3531..3550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3784..3801
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3918..3939
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3954..3959,3960..3977,3978..3984)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3992..4008
                     /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     3997..4019
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4016..4032
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4016..4032
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4081..4202
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4092..4111)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4146..4165
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4376..5170)
                     /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(4487..4506)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5097..5116
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5237..5566)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5251..5386
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5305..5324)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5551..5571
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5580..6008)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5690..5711)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5902..5921
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    6054..6278
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6267..6284
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(7872..7897,1..583,584..586,587..934))
                     /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"
ORIGIN
        1 ctccgcccat ccagggcttg atgattccgt cctgctccag cagcagagct ccctgcttgt
       61 cgattcctct ggagatgccg aagatttcct tgtcgccgat gatcagcttc acaggtctgt
      121 tgatgaagtt gtccagcttc tcccatctgc tcaggtaagg agccagtccc tcctgctcga
      181 acagctccag agcggctctc agctctctga tcagcatggc ggccagggtg ttccggtcca
      241 ggttgatgcc agcctcctgc agggtgatcc agccctggtt caccacgctc tcctccactc
      301 ttctcatggc catgttgatt ccggctccga tcacgatctg agcggcatcg cctgtcttgc
      361 ctgtcagctc caccaggatg ccagccagct tgcggtcctg gaggtacagg tcattaggcc
      421 acttcactct caccttgtcg gctcccagct ttctcagcac ctcggccatc acgattccga
      481 tcaccagact cagtccgatg gcagcagcag gtccctgctc cagtctccag aacatggaca
      541 ggtacaggtt ggctccgaat gggctgaacc acttccgtcc tctgcctcct ctgccagcct
      601 gctggtactc ggcgatgcag gcgtcgccgg acttcagctc tccgattctg tccagcaggt
      661 actgattggt ggagtcgatc acaggcagca cggccacgct tccgccatcc agctgtccca
      721 ggatctgctt ggcgttcagc agctggatag gctcaggcag gctgtagccc ttgccaggca
      781 cggtgaacac gtccactccc cagtctctca gtgtctggat gtgcttgttg atggcggctc
      841 tgctcattcc cagggtctct cccagctgct cgccagagtg gaactcgccg ttggccagca
      901 gggcgatcag cttcaggggc acggtgttgt ccttgtcgag atcctcttct gagatgagtt
      961 tttgttccat ggtggctagc cagcttgggt ctccctatag tgagtcgtat taatttcgat
     1021 aagccagtaa gcagtgggtt ctctagttag ccagagagct ctgcttatat agacctccca
     1081 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     1141 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     1201 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     1261 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     1321 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     1381 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     1441 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     1501 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     1561 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     1621 actagtcaat aatcaatgtc aacgcgtata tctggcccgt acatcgcgaa gcagcgcaaa
     1681 acgcctaacc ctaagcagat tcttcatgca attgtcggtc aagccttgcc ttgttgtagc
     1741 ttaaattttg ctcgcgcact actcagcgac ctccaacaca caagcaggga gcagatactg
     1801 gcttaactat gcggcatcag agcagattgt actgagagtg caccataggg gatcgggaga
     1861 tctcccgatc cgtcgacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1921 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1981 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2041 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2101 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2161 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2221 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     2281 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     2341 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2401 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2461 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2521 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     2581 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2641 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2701 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     2761 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2821 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2881 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2941 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3001 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3061 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3121 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3181 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3241 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3301 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3361 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3421 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3481 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3541 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3601 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3661 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3721 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3781 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     3841 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     3901 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     3961 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     4021 aacagctatg accatgatta cgccaagctc tagctagagg tcgacggtat acagacatga
     4081 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     4141 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     4201 ttggggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg
     4261 tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct
     4321 cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga
     4381 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
     4441 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
     4501 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag
     4561 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
     4621 gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc
     4681 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     4741 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     4801 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     4861 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     4921 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     4981 cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     5041 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     5101 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     5161 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agatccgaaa atggatatac
     5221 aagctcccgg gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct
     5281 ggaatagctc agaggcagag gcggcctcgg cctctgcata aataaaaaaa attagtcagc
     5341 catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg
     5401 gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
     5461 agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc
     5521 tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc cacagaatta
     5581 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     5641 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     5701 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     5761 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     5821 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     5881 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     5941 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     6001 gggcgcgtgg ggataccccc tagagcccca gctggttctt tccgcctcag aagccataga
     6061 gcccaccgca tccccagcat gcctgctatt gtcttcccaa tcctccccct tgctgtcctg
     6121 ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg caatttcctc
     6181 attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag gcacggggga
     6241 ggggcaaaca acagatggct ggcaactaga aggcacagtc gaggctgatc agcggtttaa
     6301 acttaagctt ggtaccgagc tcggatcctt agatgttttg ccacaaagag gtgcctcctg
     6361 agaagttttc tttcccattt ggaatacagg gtggcttgat gggtactgtt ggtgaccccg
     6421 agcgatgaaa atgaacattc tgccactttc catcccggcg gtgccacaca cgagtctctt
     6481 ctgactgcat tgtctttggc attccactgc catccatgta ctgtgtgagc ctaatatatg
     6541 ctatgcaggc ggcatcatcc cctaccagat gtacatgagg gtttagaata atagtgtgga
     6601 ttggtttatt gcttttggac aaagcatttt caaagtagaa tcggtgaaaa tccatccctt
     6661 ccactaaatt acccaaagct tcaggttcaa aagcagtaag gcctgggtca cagatttttg
     6721 tgtaggcttc aaagtcccca ttgttgatag cttcgatcag ttgttcagtg actttgataa
     6781 tctcttgctt tcgtgctttc acatcttcat cctcaattgt tgtatttgaa ctctcagttg
     6841 actcctttac tccatctggt ttcttcaaca aactcttggc tgctgagaaa ttccttgtag
     6901 ccagcatagt tgtcaagatg gcacccttta gttttcttct agcattaaat ttcttcaagc
     6961 agtctacagt ctcctgtctg tgcatcatgg aagcaacagt agaacgttga cagatccatg
     7021 ggtgcttcag tgcctctgag gctgtgatgc gtttggcagg gttgatagta agcattttat
     7081 tgatgaggtc tttggcttca ggagtcaccg tgtcccattc tggtgatgga aaatcataag
     7141 ctccagcctt gatctgctga tagagtctgt gttggtcttc atcccagaag ggtggatacc
     7201 ccacaagtag aatatagaga atgacaccac atgcccacat atccactggc tttccataag
     7261 gatctttacg taaaacttct ggagaaagat atccaggtgt gccagcaaaa ccaaaccacg
     7321 cctgctggtc cccttgaact tctatggcta agccaaagtc tgccaatttc acagctgctc
     7381 ccttggattt gctagctaaa agcaaattct caggcttcag gtccctgtga actatgccat
     7441 ttaggtgaca atgattaaca ctttctagaa tctgctgtat acaatgactg gcatcagctt
     7501 cactgtagta ttctcttgcc actatgtctt caaacagttc acctccagta actaaatcaa
     7561 acaccaagta gtgaaagccc tcttctgata tgctatcatg aagtcgcaca atattagggt
     7621 gcttcaaaag acggcagatt ctagcttctc tttctagttt ctgatgatcc ctagcagaaa
     7681 gctttttggt gttgataatt ttggcagcat attcttgtcc agtaggaatt ttcatacatc
     7741 ttctcaccac tgagaatgcc ccctttccaa gctcctcgaa aagctgatac tcgtccgtga
     7801 acctggtgca ggttgtggtc gaagccatga attctttctg cagatatcca gcacagtggc
     7861 ggccgctcga gcttctctgc gcttctcagg gagattt
//
