LOCUS       sequence-200662-        7173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  heat shock inducible transgenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7173)
  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 7173)
  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-DN-Cep152-mKate2
FEATURES             Location/Qualifiers
     source          1..7173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..22
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3352..4050
                     /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    4177..4298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4214..4233)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4268..4287
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4305..4760)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4442..4463)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4654..4673
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4787..4891
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4893..5250
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4893..5084
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4916..4936)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5101..5236
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5285..6079
                     /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             5285..6079
                     /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(5339..5358)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5949..5968
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6267..6286)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6311..6358
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(6687..7173,1..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 gcagtcatgt cattagactt tggcagtgtg gcactaccag tgcaaaatga agatgaagag
     1861 tatgacgaag aggactatga aagagagaaa gagttgcagc agttactcac agaccttccc
     1921 catgacatgc tggatgacga cctctcctct ccagagctcc agtattcgga ctgcagcgag
     1981 gatggcacag acggacaacc acatcatcct gagcaattgg agatgagctg gaatgagcaa
     2041 atgctgccca aatctcaaag tgtaaatggc tataatgaaa ttcagagttt atatgctgga
     2101 gaaaaatgtg gtaatgtctg ggaagaaaat agaagtaaaa ctgaagaccg acatcctgtg
     2161 taccatcctg aagaaggtgg agatgaaggt ggaagtggtt atagtcctcc aagtaaatgt
     2221 gaacagactg atttatatca ccttcctgaa aactttaggc catataccaa tggtcagaag
     2281 caggaattta ataaccaagc aaccaatgta attaaatttt cagatcctca atggaaccat
     2341 tttcagggtc ccagttgtca aggtttggaa ccgtataata aagtgacata taaaccttat
     2401 cagtcttctg cccagaataa tggctcacca gcccaggaga taacaggaag tgacacattc
     2461 gaaggcctgc aacaacaatt tttaggagct aatgaaaact ctgcagaaaa tatgcagatt
     2521 attcaacttc aggttcttaa caaagcaaaa gagagacaac tggagaactt aattgaaaag
     2581 ttaaatgaaa gtgaacgtca aattcgatat ctgaatcacc agcttgtaat aataaaagat
     2641 gaaaaggatg gtttgactct cagccttcga gaatcacaga aactctttca gaatggaaaa
     2701 gaaagagaga tacagcttga agctcaaata aaagcactgg agactcagat acaagcatta
     2761 aaagtcaatg aagaacagat gatcaagaag tccagaacaa ctgaaatggc tctggaaagc
     2821 ttgaagcagc agctggtgga ccttcatcat tctgaatcac ttcaacgagc tagagaacag
     2881 catgagagca ttgttatggg cctcacaaag aagtacgaag agcaagtatt gtccttacaa
     2941 aagaatttgg atgccacagt caccgcactt aaagaacagg aagacatttg ctctcgtctg
     3001 aaagatcacg tgaaacaact ggaaaggaat caagaagcaa tcaagttaga aaagactgag
     3061 atcattaata agttgacaag aagtctagag gagagtcaaa agcagtgtgc ccacttgttg
     3121 cagtccgggt cagtacaaga ggtggctcag ctacagttcc agctgcagca agcacagaag
     3181 gcacatgcta tgagtgcaaa catgaacaag gctttgcaag aagaattaac agaactaaaa
     3241 gatgaaattt ctctctatga atctgctgca aaactaggaa tacatccaag tgactcagaa
     3301 ggagaattaa atatagaact cactgaatcg tatgtggatt tggtaggatc catggtgagc
     3361 gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt gaacaaccac
     3421 cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga
     3481 atcaaggcgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc taccagcttc
     3541 atgtacggca gcaaaacctt catcaaccac acccagggca tccccgactt ctttaagcag
     3601 tccttccccg agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg
     3661 accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga
     3721 ggggtgaact tcccatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc
     3781 tccaccgaga ccctgtaccc cgctgacggc ggcctggaag gcagagccga catggccctg
     3841 aagctcgtgg gcgggggcca cctgatctgc aacttgaaga ccacatacag atccaagaaa
     3901 cccgctaaga acctcaagat gcccggcgtc tactatgtgg acagaagact ggaaagaatc
     3961 aaggaggccg acaaagagac ctacgtcgag cagcacgagg tggctgtggc cagatactgc
     4021 gacctcccta gcaaactggg gcacagatga cccgggcggc cgcgactcta gatcataatc
     4081 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     4141 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     4201 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4261 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     4321 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4381 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4441 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     4501 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     4561 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     4621 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     4681 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     4741 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4801 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4861 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     4921 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4981 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5041 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5101 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5161 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     5221 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     5281 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     5341 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5401 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     5461 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5521 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5581 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5641 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5701 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5761 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     5821 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5881 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5941 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     6001 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     6061 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     6121 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     6181 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     6241 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     6301 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     6361 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     6421 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     6481 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     6541 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6601 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6661 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6721 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6781 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     6841 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6901 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6961 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7021 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7081 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7141 gtcggaacag gagagcgcac gagggagctt cca
//
