LOCUS       sequence-203909-        8441 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 8441)
  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 8441)
  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-Ras-mKate2
FEATURES             Location/Qualifiers
     source          1..8441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(34..462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     356..375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     589..611
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     603..620
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     604..620
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     630..649
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        630..648
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(778..789,790..952)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     protein_bind    1324..1344
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    2864..2888
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2891..3586
                     /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"
     protein_bind    complement(3666..3690)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3695..3829
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3719..3738)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3773..3792
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(5971..6158,6159..6170)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(6204..6220)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6204..6220)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(6257..6277)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6257..6275)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6296..6312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6296..6312)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6309..6331)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6320..6336
                     /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(6344..6350,6351..6368,6369..6374))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6389..6410
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6527..6544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6698..7286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6778..6797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7457..8248,8249..8317))
                     /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     8080..8099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8318..8422)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      241 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      301 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      361 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
      421 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
      481 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
      541 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
      601 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
      661 caggtcattg cacaacacca gaaatgccct ctgatctgca aaagacgtga atatctgttc
      721 agacacccat atccactctg ttccacacag gtcagaggtt tgtccaggag ttcttgacag
      781 aggtgtaaaa agtactcaaa aattttactc aagtgaaagt acaagtactt agggaaaatt
      841 ttactcaatt aaaagtaaaa gtatctggct agaatcttac ttgagtaaaa gtaaaaaagt
      901 actccattaa aattgtactt gagtattaag gaagtaaaag taaaagcaag aaagaaaact
      961 agagattctt gtttaagctt ttaatctcaa aaaacattaa atgaaatgca tacaaggttt
     1021 tatcctgctt tagaactgtt tgtatttaat tatcaaacta taagacagac aatctaatgc
     1081 cagtacacgc tactcaaagt tgtaaaacct cagatttaac ttcagtagaa gctgattctc
     1141 aaaattgtta gtgtcaagcc tagctctttt ggggctgaaa agcaatcctg cagtgctgaa
     1201 aagcctctca caggcagccg atgcgggaag aggtgtatta gtcttgatag agaggctgca
     1261 aatagcagga aacgtgagca gagactccct ggtgtctgaa acacaggcca gatgggccga
     1321 tatcaacttt gtatagaaaa gttgcttcag gggtgtcgct tggttatttc caaaaatcaa
     1381 attaatttta ttaaactatt agaacgagca tgttttgtct atatgctaca gaagataaaa
     1441 aataatagga gttaacagtt ataaaacaac acactttgtt tctattgatt gttgaccaca
     1501 ctggggtctc attaagttag attaaagaca cactaactgg gtcaaaagca gcagattgat
     1561 ttcatagcac cagggtaaac tttctaacac ttttacggca atcatataca ttaaaattaa
     1621 atacagacca cgactgaaca aggaggatga tctccaatat taaacaaaga gacttgtgcc
     1681 tatttctctg agggtaaaca tgacctctca agttagcaag ttggttttaa cactacaaaa
     1741 atagttaaga ctgcaatccc agaataaagt attggtttta accaatcaat atagtacagt
     1801 aaacatccat ttgttttgtt gaaacgttaa acaaatctga ccaaagctat tagcttatat
     1861 aaaacaggtt tgccttctat gtagctgaaa acaccacagg cccgattttg ctactgtgta
     1921 aaacatttca gcaagatttt tttattgcat ttttttttac tgaatcgttc aaacatttta
     1981 tcattttagt ttgttcattc attgcaactg gaaaaacaac acatcacaca accgcacata
     2041 tttcagcaat aagtacaata aaacactcaa ataaaaaaaa cattttaaat ctctttgtat
     2101 ttttgaccgc tgtttcgcgt aatttcacgg taaaactctg gaaatctcca ctacattcct
     2161 ctcagcggct cctctcaatg acagctgaag aagtgacgcg gctgcctgct gtgttttgat
     2221 tggtcgaatt cactggaggc ttccagaaca gtgtagagtc tgaacgggtg cgcgctctgc
     2281 tgtatttaaa gggcgaaaga gagaccgcag agaaactcaa ccgaagagaa gcgacttgac
     2341 aaagaagaaa agagcagcct gacaggactt tttccccgac gaggtgttta ttcgctctat
     2401 ttaagaatct actgtaaggt aagtctcaat atattgtact ctattggcta atcagaatta
     2461 tatagagatt atatgtactt aatgtcaaaa aatcaacttt gtatatgtaa tctttttaca
     2521 tgtggactgc ctatgttcat cttattttag gtctactaga aaattatatt tcccgttttc
     2581 acaataagga tttttaaaaa aagcaatgaa cagacgggca tttactttat gttgctgaca
     2641 ttattttata tgagcataat aaccataaat actagcaaat gtcctaaatg aatttgtgtt
     2701 aatgttgtct acaaaagaaa attagcgttt tacttgtaca actaataata acttggttat
     2761 taagagaatt tcacttgttg actagaaaaa tcctttcata atgaaacaat tgcaccataa
     2821 attgtataaa tataaaatta attctaattg tttttttttc ctgccaagtt tgtacaaaaa
     2881 agcaggctgg atggtgagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga
     2941 gggcaccgtg aacaaccacc acttcaagtg cacatccgag ggcgaaggca agccctacga
     3001 gggcacccag accatgagaa tcaaggcggt cgagggcggc cctctcccct tcgccttcga
     3061 catcctggct accagcttca tgtacggcag caaaaccttc atcaaccaca cccagggcat
     3121 ccccgacttc tttaagcagt ccttccccga gggcttcaca tgggagagag tcaccacata
     3181 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
     3241 ctacaacgtc aagatcagag gggtgaactt cccatccaac ggccctgtga tgcagaagaa
     3301 aacactcggc tgggaggcct ccaccgagac cctgtacccc gctgacggcg gcctggaagg
     3361 cagagccgac atggccctga agctcgtggg cgggggccac ctgatctgca acttgaagac
     3421 cacatacaga tccaagaaac ccgctaagaa cctcaagatg cccggcgtct actatgtgga
     3481 cagaagactg gaaagaatca aggaggccga caaagagacc tacgtcgagc agcacgaggt
     3541 ggctgtggcc agatactgcg acctccctag caaactgggg cacagaagat ctcgtggatc
     3601 taagctgaac cctcctgatg agagtggccc cggctgcatg agctgcaagt gtgtgctctc
     3661 ctgagaccca gctttcttgt acaaagttga tgtagatcca gacatgataa gatacattga
     3721 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3781 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3841 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc
     3901 cgcggcgcca atgcattggg ccctcgagca tatgagatct gtcccgatct ttctcttctg
     3961 tgctgtcaat ttagagtagt ttttgaggta atttgggtgc attctctgta aaaacaacaa
     4021 ataaatattt atattgtatg caataaatgt aacaattaca catacactct ctgattacag
     4081 cattaaaggg ttagttcacc caaaaatgaa aagatgtcat taatgactca ccctcatgtc
     4141 gttccaagcc cgtaacacct ccgttcatct tcggaacaca gtttaagata ttttagattt
     4201 agtccgagag ctttctgtcc ctccactgaa aatgtatgta cggtatactg tccatgtcca
     4261 gaaaggtaat aaaaacatca aagtagtcca tgtgacatca gtgggttagg tagaattatt
     4321 tgaagcatcg aaaatacatt ttggtccaaa aataacaaaa cctacgactt tattcagcat
     4381 tgtagcgtca ctgcgaagtc gtgaacgcgg attgacaaca gacccggaag agaatacaat
     4441 gccgaataaa gtcgtaggtt ttgttatttt tggaccaaaa tgtattcgat gcttcaaaaa
     4501 attctaacta acccactgat gtcacatgga ctactttgat gtttttatta cctttctgga
     4561 catggacagt ataccgtaca tacattctca atggaggcac agaaagctct cggactaaat
     4621 ctaaaatatc ttaaactgtg ttctgaagat gaacggaggt cttacgggct tggaacgaca
     4681 tgagggcgag tcattaatga cattattttc atttttgggt gaactaaccc tttaataaaa
     4741 ctgacatttg cagccaattg cacactttat aatgttaaat acttgttgat tttagaaata
     4801 cagtagcatt tagtgacaat caaacattaa ttttatttct gattatattg acaaaaacac
     4861 aggactgtaa ggctgacatg acagggtgaa ttgatttcat tacattagtt ttgatggtaa
     4921 cacttttaca ataaggttcg ttagttaact acattagtta acatgaacta ataatgaact
     4981 gcacgtatac atgttaattt caacatttac taatacttta tatcttgtta acattagtta
     5041 atgcactgtg aactaacatg aacaaacaat gaacatctgt atttctatta actaacgtta
     5101 acaaagatta acagatacag taacaaatgt attgctcatg gttatttaat gttagttaat
     5161 acatgaacta atcaacctca ttgtaaagtg ttaccgtttt gacaatacaa catgtttgga
     5221 aaaaaatttc tcgcttggtt ttcttgtgtt ttgcatgaat tcaaacacaa tgagagcacg
     5281 gaatgataat gattattcaa agctgcaata ggcgccaaat ttcccgtgga cctaacgtta
     5341 ccaattacaa atgcatttta gcattttatt acgtgagcct tatattgtca atcgcatgca
     5401 ttagtgaaaa taggcctact acacatataa aaggcgcgcc atcaacgtca aaacatgcac
     5461 acccaaaaaa aaaaaaaaaa aaaaaaaaaa aaanctatca gtaaaacaaa atacttaatg
     5521 tacttacctc aatatgcttc cttaggtttg atggcgaact tttgaaggcc gatatttctt
     5581 tattaagcgg gagacagagg acacatttca tcttgaatga atctttattt acaccactta
     5641 aagaaaagaa ttcgcgaaga tacggccacg ggtgctcttg atcctgtggc tgattttgga
     5701 ctgtgctgct cgcagctgct gatgaatcac atacttcctc cattttcttc cactgattga
     5761 ctgttataat ttccctaatt tccaggtcaa ggtgctgtgc attgtggtaa tagatgtgac
     5821 atgacgtcac ttccaaagga ccaatgaaca tgtctgacca atttcatata atgtgaaaac
     5881 gattttcata ggcagaataa ataacattta aattaaactg ggcatcagcg caattcaatt
     5941 ggtttggtaa tagcaaggga aaatagaatg aagtgatctc caaaaaataa gtactttttg
     6001 actgtaaata aaattgtaag gagtaaaaag tacttttttt tctaaaaaaa tgtaattaag
     6061 taaaagtaaa agtattgatt tttaattgta ctcaagtaaa gtaaaaatcc ccaaaaataa
     6121 tacttaagta cagtaatcaa gtaaaattac tcaagtactt tacacctctg gttcttgacc
     6181 ccctaccttc agcaagccca gcagatccac tagttctaga gcggccgcca ccgcggtgga
     6241 gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     6301 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6361 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6421 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6481 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6541 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6601 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6661 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6721 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6781 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6841 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6901 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6961 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7021 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7081 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     7141 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7201 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7261 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7321 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7381 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7441 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7501 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7561 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7621 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7681 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7741 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7801 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7861 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7921 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7981 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8041 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8101 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8161 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8221 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8281 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8341 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8401 ataaacaaat aggggttccg cgcacatttc cccgaaaagt g
//
