LOCUS       sequence-200664-        8352 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  heat shock inducible transgenesis, non phoshorylable version of 
            Plk4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8352)
  AUTHORS   Strzyz PJ, Lee HO, Sidhaye J, Weber IP, Leung LC, Norden C
  TITLE     Interkinetic nuclear migration is centrosome independent and ensures
            apical cell division to maintain tissue integrity.
  JOURNAL   Dev Cell. 2015 Jan 26;32(2):203-19. doi: 
            10.1016/j.devcel.2014.12.001. Epub 2015 Jan 15.
  PUBMED    25600237
REFERENCE   2  (bases 1 to 8352)
  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: 
            "10.1016/j.devcel.2014.12.001"; journalName: "Dev Cell"; date: 
            "2015-01-26- 26"; volume: "32"; issue: "2"; pages: "203-19"
COMMENT     Sequence Label: HS-OS-Plk4-mKate2
FEATURES             Location/Qualifiers
     source          1..8352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..22
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             4529..5227
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLXRIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     polyA_signal    5356..5477
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5393..5412)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5447..5466
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5484..5939)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5621..5642)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5833..5852
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5966..6070
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6072..6429
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6072..6263
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6095..6115)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6280..6415
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6464..7258
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6464..7258
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6518..6537)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7128..7147
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7446..7465)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7490..7537
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(7866..8352,1..102)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
       61 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      121 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
      181 cctgattctg tggataaccg tattaccgcc atgcattagt tattactagc gctaccggac
      241 tcagatctcg agctcaagct tcgaattctg cagcccgggg gatccttcag gggtgtcgct
      301 tggttatttc caaaaatcaa attaatttta ttaaactatt agaacgagca tgttttgtct
      361 atatgctaca gaagataaaa aataatagga gttaacagtt ataaaacaac acactttgtt
      421 tctattgatt gttgaccaca ctggggtctc attaagttag attaaagaca cactaactgg
      481 gtcaaaagca gcagattgat ttcatagcac cagggtaaac tttctaacac ttttacggca
      541 atcatataca ttaaaattaa atacagacca cgactgaaca aggaggatga tctccaatat
      601 taaacaaaga gacttgtgcc tatttctctg agggtaaaca tgacctctca agttagcaag
      661 ttgtttttaa cactacaaaa atagttaaga ctgcaatccc agaataaagt attggtttta
      721 accaatcaat atagtacagt aaacatccat ttgttttgtt gaaacgttaa acaaatctga
      781 ccaaagctat tagcttatat aaaacaggtt tgccttctat gtagctgaaa acaccacagg
      841 cccgattttg ctactgtgta aaacatttca gcaagatttt tttattgcat tttttttact
      901 gaatcntcaa acattttatc attttagttt gttcattcat tgcaactgga aaaacaacac
      961 atcacacaac cgcacatatt tcagcaataa gtacaataaa acactcaaat aaaaaaaaca
     1021 ttttaaatct ctttgtattt ttgaccgctg tttcgcgtaa tttcacggta aaactctgga
     1081 aatctccact acattcctct cagcggctcc tctcaatgac agctgaagaa gtgacgcggc
     1141 tgcctgctgt gttttgattg gtcgaattca ctggaggctt ccagaacagt gtagagtctg
     1201 aacgggtgcg cgctctgctg tatttaaagg gcgaaagaga gaccgcagag aaactcaacc
     1261 gaagagaagc gacttgacaa agaagaaaag agcagcctga caggactttt tccccgacga
     1321 ggtgtttatt cgctctattt aagaatctac tgtaaggtaa gtctcaatat attgtactct
     1381 attggctaat cagaattata tagagattat atgtacttaa tgtcaaaaaa tcaactttgt
     1441 atatgtaatc tttttacatg tggactgcct atgttcatct tattttaggt ctactagaaa
     1501 attatatttc ccgttttcac aataaggatt tttaaaaaaa gcaatgaaca gacgggcatt
     1561 tactttatgt tgctgacatt attttatatg agcataataa ccataaatac tagcaaatgt
     1621 cctaaatgaa tttgtgttaa tgttgtctac aaaagaaaat tagcgtttta cttgtacaac
     1681 taataataac ttggttatta agagaatttc acttgttgac tagaaaaatc ctttcataat
     1741 gaaacaattg caccataaat tgtataaata taaaattaat tctaattgtt tttttttcct
     1801 gcagtcgatg gcggcgtgca tcggggagag gatcgaggac tttaaggttg gaaatctact
     1861 cggtaaagga tcatttgctg gtgtctacag agctgagtcc atacacactg gtttggaagt
     1921 tgcaatcaaa atgatagata agaaagccat gtacaaagct ggaatggtac agagagtcca
     1981 aaatgaggtg aaaatacatt gccagttgaa acacccctct gtcttggagc tctataatta
     2041 ctttgaagat aacaattatg tctacctggt attggaaatg tgccacaatg gagaaatgaa
     2101 cagatatctg aagaacagaa tgaagccttt ctcagaaagg gaagctaggc acttcatgca
     2161 ccagattatc acaggaatgt tatatcttca ttctcatggc atattgcacc gggacctcac
     2221 actctctaac atcttactta cgcggaatat gaacataaaa attgctgact ttggactagc
     2281 aacgcagttg aatatgccac atgaaaagca ctatacactc tgtgggactc ctaattatat
     2341 ttcaccagaa attgcaactc gaagtgcaca tggacttgaa tctgatattt ggtcattggg
     2401 ctgtatgttt tatacgttac ttattggaag accacctttt gacactgaca cagtcaagaa
     2461 cacattgaac aaagtagtcc tggcagatta tgaaatgcca gcctttttgt cacgagaggc
     2521 ccaggacctt atccaccagt tacttcgtag aaaccctgca gatcggttaa gtctgtcttc
     2581 tgtgttggac catcctttca tgtcacgaaa tccttcacca aagagtaaag acgtagggac
     2641 tgtagaggac ctacttgaca gaagactttt ggttggtcaa ccacttccaa ataaaattac
     2701 tgtatttcaa aaaaataaaa attcaagtga cttttcttca ggagatggaa gtaatttttg
     2761 tactcaatgg ggaaatccag aacaagaagc taatagtagg ggacggggga gagtgattga
     2821 agatgcagaa gagaggccgc attctcgata cctgcgcaga gctcattcct ctgatagagc
     2881 cagcccctct aatcagtctc gagccaaaac atactcagta gaacgttgtc actcagtaga
     2941 aatgctttca aagcctagaa gatcactgga tgaaaatcaa cacagttcca atcatcattg
     3001 tctaggaaaa actccttttc catttgcaga ccagacacct cagatggaaa tggtacagca
     3061 gtggtttggg aatctgcaaa tgaatgctca tttaggagaa actaatgagc accacaccgt
     3121 tagcccaaac agagatttcc aggactatcc agatttgcag gacacgttac gaaacgcttg
     3181 gactgacacg agagccagca agaatgctga tacttctgcc aatgttcatg ctgtaaagca
     3241 gctgagtgcc atgaaataca tgagtgcaca tcaccataag cctgaggtca tgccacagga
     3301 gccggaccta catcctcatt ctgaacaaag caagaataga agtatggagt cgacactggg
     3361 ttaccagaaa cctaccttaa gaagtattac atctcctctg attgctcaca gattaaagcc
     3421 aatcagacag aaaaccaaaa aggctgtggt gagcatcctt gattcagagg aggtgtgtgt
     3481 ggagcttctg agagagtgtg agtctgaagg atatgtgaaa gaagtgcttc agatatcgag
     3541 tgatgggact atgatcactg tttattaccc gaacgatgga agaggctttc ctcttgctga
     3601 cagacctccc ttgcctacta acaacatcag taggtacagc tttgacaatc taccagaaaa
     3661 atactggcgg aaatatcagt atgcttccag attcattcag ctagtaagat ctaaaactcc
     3721 caaaatcact tattttacaa gatatgctaa atgtattttg atggaaaatt ctcctggtgc
     3781 tgatttcgaa gtttggtttt atgatggagc caaaatacat aaaactgaaa atttaattca
     3841 cataattgag aaaacaggga tatcttataa tttaaaaaat gaaaatgaag ttaccagcct
     3901 gaaagaggaa gtaaaagtat atatggacca tgctaatgag ggtcaccgta tttgcttgtc
     3961 actggaatct gtaatctctg aggaggaaaa gagaagcagg ggttcttcat tcttccctat
     4021 aatcgtagga agaaaacctg gtaatactag ttcacctaaa gccttatcag ctcctcctgt
     4081 ggacccaagc tgctgtaagg gagagcaggc gtcagcaagc agactgagcg tgaatagtgc
     4141 cgctttcccc acacagtccc caggactcag tccttccact gtgacagttg aaggacttgg
     4201 ccacacagcg actgccacag gaacaggcgt ctcttcaagt cttcctaaat ctgcacagct
     4261 tttgaaatct gtttttgtga aaaatgttgg ttgggctaca cagctaacta gcggagctgt
     4321 gtgggttcag tttaatgatg ggtcacagtt ggttgtccag gcaggagtat cttccatcag
     4381 ttacacatca ccagatggtc agacaactag gtatggagaa aatgaaaaat tacctgaata
     4441 catcaaacag aaattacagt gtctttcttc catccttctg atgttttcta atccaactcc
     4501 taattttcag accggttcat cgggatccat ggtgagcgag ctgattaagg agaacatgca
     4561 catgaagctg tacatggagg gcaccgtgaa caaccaccac ttcaagtgca catccgaggg
     4621 cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggcggtcg agggcggccc
     4681 tctccccttc gccttcgaca tcctggctac cagcttcatg tacggcagca aaaccttcat
     4741 caaccacacc cagggcatcc ccgacttctt taagcagtcc ttccccgagg gcttcacatg
     4801 ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct
     4861 ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc catccaacgg
     4921 ccctgtgatg cagaagaaaa cactcggctg ggaggcctcc accgagaccc tgtaccccgc
     4981 tgacggcggc ctggaaggca gagccgacat ggccctgaag ctcgtgggcg ggggccacct
     5041 gatctgcaac ttgaagacca catacagatc caagaaaccc gctaagaacc tcaagatgcc
     5101 cggcgtctac tatgtggaca gaagactggn aagaatcaag gaggccgaca aagagaccta
     5161 cgtcgagcag cacgaggtgg ctgtggccag atactgcgac ctccctagca aactggggca
     5221 cagatgagac ccgggcggcc gcgactctag atcataatca gccataccac atttgtagag
     5281 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     5341 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5401 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5461 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     5521 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     5581 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     5641 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     5701 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     5761 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     5821 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     5881 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     5941 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6001 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6061 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     6121 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     6181 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     6241 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     6301 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6361 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     6421 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     6481 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     6541 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     6601 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     6661 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     6721 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     6781 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     6841 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     6901 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     6961 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     7021 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     7081 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     7141 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     7201 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     7261 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     7321 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     7381 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     7441 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     7501 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     7561 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     7621 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     7681 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     7741 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7801 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7861 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7921 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7981 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     8041 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     8101 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     8161 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     8221 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     8281 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     8341 agggagcttc ca
//
