LOCUS       sequence-203461-        8353 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 8353)
  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 8353)
  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..8353
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..22
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             4530..5228
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     polyA_signal    5357..5478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5394..5413)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5448..5467
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5485..5940)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5622..5643)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5834..5853
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5967..6071
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6073..6430
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6073..6264
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6096..6116)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6281..6416
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6465..7259
                     /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             6465..7259
                     /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(6519..6538)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7129..7148
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7447..7466)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7491..7538
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(7867..8353,1..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 ttttttttac
      901 tgaatcgttc aaacatttta tcattttagt ttgttcattc attgcaactg gaaaaacaac
      961 acatcacaca accgcacata tttcagcaat aagtacaata aaacactcaa ataaaaaaaa
     1021 cattttaaat ctctttgtat ttttgaccgc tgtttcgcgt aatttcacgg taaaactctg
     1081 gaaatctcca ctacattcct ctcagcggct cctctcaatg acagctgaag aagtgacgcg
     1141 gctgcctgct gtgttttgat tggtcgaatt cactggaggc ttccagaaca gtgtagagtc
     1201 tgaacgggtg cgcgctctgc tgtatttaaa gggcgaaaga gagaccgcag agaaactcaa
     1261 ccgaagagaa gcgacttgac aaagaagaaa agagcagcct gacaggactt tttccccgac
     1321 gaggtgttta ttcgctctat ttaagaatct actgtaaggt aagtctcaat atattgtact
     1381 ctattggcta atcagaatta tatagagatt atatgtactt aatgtcaaaa aatcaacttt
     1441 gtatatgtaa tctttttaca tgtggactgc ctatgttcat cttattttag gtctactaga
     1501 aaattatatt tcccgttttc acaataagga tttttaaaaa aagcaatgaa cagacgggca
     1561 tttactttat gttgctgaca ttattttata tgagcataat aaccataaat actagcaaat
     1621 gtcctaaatg aatttgtgtt aatgttgtct acaaaagaaa attagcgttt tacttgtaca
     1681 actaataata acttggttat taagagaatt tcacttgttg actagaaaaa tcctttcata
     1741 atgaaacaat tgcaccataa attgtataaa tataaaatta attctaattg tttttttttc
     1801 ctgcagtcat ggcggcgtgc atcggggaga ggatcgagga ctttaaggtt ggaaatctac
     1861 tcggtaaagg atcatttgct ggtgtctaca gagctgagtc catacacact ggtttggaag
     1921 ttgcaatcaa aatgatagat aagaaagcca tgtacaaagc tggaatggta cagagagtcc
     1981 aaaatgaggt gaaaatacat tgccagttga aacacccctc tgtcttggag ctctataatt
     2041 actttgaaga taacaattat gtctacctgg tattggaaat gtgccacaat ggagaaatga
     2101 acagatatct gaagaacaga atgaagcctt tctcagaaag ggaagctagg cacttcatgc
     2161 accagattat cacaggaatg ttatatcttc attctcatgg catattgcac cgggacctca
     2221 cactctctaa catcttactt acgcggaata tgaacataaa aattgctgac tttggactag
     2281 caacgcagtt gaatatgcca catgaaaagc actatacact ctgtgggact cctaattata
     2341 tttcaccaga aattgcaact cgaagtgcac atggacttga atctgatatt tggtcattgg
     2401 gctgtatgtt ttatacgtta cttattggaa gaccaccttt tgacactgac acagtcaaga
     2461 acacattgaa caaagtagtc ctggcagatt atgaaatgcc agcctttttg tcacgagagg
     2521 cccaggacct tatccaccag ttacttcgta gaaaccctgc agatcggtta agtctgtctt
     2581 ctgtgttgga ccatcctttc atgtcacgaa atccttcacc aaagagtaaa gacgtaggga
     2641 ctgtagagga cctacttgac agaagacttt tggttggtca accacttcca aataaaatta
     2701 ctgtatttca aaaaaataaa aattcaagtg acttttcttc aggagatgga agtaattttt
     2761 gtactcaatg gggaaatcca gaacaagaag ctaatagtag gggacggggg agagtgattg
     2821 aagatgcaga agagaggccg cattctcgat acctgcgcag agctcattcc tctgatagag
     2881 ccagcccctc taatcagtct cgagccaaaa catactcagt agaacgttgt cactcagtag
     2941 aaatgctttc aaagcctaga agatcactgg atgaaaatca acacagttcc aatcatcatt
     3001 gtctaggaaa aactcctttt ccatttgcag accagacacc tcagatggaa atggtacagc
     3061 agtggtttgg gaatctgcaa atgaatgctc atttaggaga aactaatgag caccacaccg
     3121 ttagcccaaa cagagatttc caggactatc cagatttgca ggacacgtta cgaaacgctt
     3181 ggactgacac gagagccagc aagaatgctg atacttctgc caatgttcat gctgtaaagc
     3241 agctgagtgc catgaaatac atgagtgcac atcaccataa gcctgaggtc atgccacagg
     3301 agccggacct acatcctcat tctgaacaaa gcaagaatag aagtatggag tcgacactgg
     3361 gttaccagaa acctacctta agaagtatta catctcctct gattgctcac agattaaagc
     3421 caatcagaca gaaaaccaaa aaggctgtgg tgagcatcct tgattcagag gaggtgtgtg
     3481 tggagcttct gagagagtgt gagtctgaag gatatgtgaa agaagtgctt cagatatcga
     3541 gtgatgggac tatgatcact gtttattacc cgaacgatgg aagaggcttt cctcttgctg
     3601 acagacctcc cttgcctact aacaacatca gtaggtacag ctttgacaat ctaccagaaa
     3661 aatactggcg gaaatatcag tatgcttcca gattcattca gctagtaaga tctaaaactc
     3721 ccaaaatcac ttattttaca agatatgcta aatgtatttt gatggaaaat tctcctggtg
     3781 ctgatttcga agtttggttt tatgatggag ccaaaataca taaaactgaa aatttaattc
     3841 acataattga gaaaacaggg atatcttata atttaaaaaa tgaaaatgaa gttaccagcc
     3901 tgaaagagga agtaaaagta tatatggacc atgctaatga gggtcaccgt atttgcttgt
     3961 cactggaatc tgtaatctct gaggaggaaa agagaagcag gggttcttca ttcttcccta
     4021 taatcgtagg aagaaaacct ggtaatacta gttcacctaa agccttatca gctcctcctg
     4081 tggacccaag ctgctgtaag ggagagcagg cgtcagcaag cagactgagc gtgaatagtg
     4141 ccgctttccc cacacagtcc ccaggactca gtccttccac tgtgacagtt gaaggacttg
     4201 gccacacagc gactgccaca ggaacaggcg tctcttcaag tcttcctaaa tctgcacagc
     4261 ttttgaaatc tgtttttgtg aaaaatgttg gttgggctac acagctaact agcggagctg
     4321 tgtgggttca gtttaatgat gggtcacagt tggttgtcca ggcaggagta tcttccatca
     4381 gttacacatc accagatggt cagacaacta ggtatggaga aaatgaaaaa ttacctgaat
     4441 acatcaaaca gaaattacag tgtctttctt ccatccttct gatgttttct aatccaactc
     4501 ctaattttca gaccggttca tcgggatcca tggtgagcga gctgattaag gagaacatgc
     4561 acatgaagct gtacatggag ggcaccgtga acaaccacca cttcaagtgc acatccgagg
     4621 gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggcggtc gagggcggcc
     4681 ctctcccctt cgccttcgac atcctggcta ccagcttcat gtacggcagc aaaaccttca
     4741 tcaaccacac ccagggcatc cccgacttct ttaagcagtc cttccccgag ggcttcacat
     4801 gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc
     4861 tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc ccatccaacg
     4921 gccctgtgat gcagaagaaa acactcggct gggaggcctc caccgagacc ctgtaccccg
     4981 ctgacggcgg cctggaaggc agagccgaca tggccctgaa gctcgtgggc gggggccacc
     5041 tgatctgcaa cttgaagacc acatacagat ccaagaaacc cgctaagaac ctcaagatgc
     5101 ccggcgtcta ctatgtggac agaagactgg aaagaatcaa ggaggccgac aaagagacct
     5161 acgtcgagca gcacgaggtg gctgtggcca gatactgcga cctccctagc aaactggggc
     5221 acagatgaga cccgggcggc cgcgactcta gatcataatc agccatacca catttgtaga
     5281 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     5341 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5401 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5461 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     5521 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     5581 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     5641 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     5701 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     5761 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     5821 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     5881 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     5941 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6001 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6061 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     6121 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6181 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6241 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     6301 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6361 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     6421 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     6481 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     6541 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     6601 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     6661 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     6721 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     6781 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     6841 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     6901 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     6961 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     7021 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     7081 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     7141 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     7201 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     7261 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     7321 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     7381 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     7441 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     7501 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     7561 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     7621 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     7681 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     7741 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7801 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7861 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7921 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7981 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     8041 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     8101 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     8161 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     8221 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     8281 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     8341 gagggagctt cca
//
