LOCUS       sequence-203029-        9226 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 9226)
  AUTHORS   Icha J, Kunath C, Rocha-Martins M, Norden C
  TITLE     Independent modes of ganglion cell translocation ensure correct 
            lamination of the zebrafish retina.
  JOURNAL   J Cell Biol. 2016 Oct 24;215(2):259-275.
  PUBMED    27810916
REFERENCE   2  (bases 1 to 9226)
  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"; journalName: "J Cell 
            Biol."; date: "2016-10-24"; volume: "215(2)"; pages: "259-275"
COMMENT     Sequence Label: hsp70:EB3-mKate2
FEATURES             Location/Qualifiers
     source          1..9226
                     /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             3752..4450
                     /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(4452..4476)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4481..4615
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4505..4524)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4559..4578
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(6756..6943,6944..6955)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(6989..7005)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6989..7005)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(7042..7062)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7042..7060)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7081..7097)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7081..7097)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7094..7116)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7105..7121
                     /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(7129..7135,7136..7153,7154..7159))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7174..7195
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7312..7329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7483..8071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7563..7582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8242..9033,9034..9102))
                     /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     8865..8884
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9103..9207)
                     /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 atggccgtca atgtgtactc cacatctgtg accagtgaaa atctgagtcg
     2941 ccatgatatg cttgcatggg tcaacgactc cctgcacctc aactatacca agatagaaca
     3001 gctttgttca ggggcagcct actgccagtt catggacatg ctcttccccg gctgtgtgca
     3061 cttgaggaaa gtgaagttcc aggccaaact agagcatgaa tacatccaca acttcaaggt
     3121 gctgcaagca gctttcaaga agatgggtgt tgacaaaatc attcctgtag agaaattagt
     3181 gaaaggaaaa ttccaagata attttgagtt tattcagtgg tttaagaaat tctttgacgc
     3241 aaactatgat ggaaaggatt acaaccctct gctggcgcgg cagggccagg acgtagcgcc
     3301 acctcctaac ccaggtgatc agatcttcaa caaatccaag aaactcattg gcacagcagt
     3361 tccacagagg acgtccccca caggcccaaa aaacatgcag acctctggcc ggctgagcaa
     3421 tgtggccccc ccctgcattc tccggaagaa tcctccatca gcccgaaatg gcggccatga
     3481 gactgatgcc caaattcttg aactcaacca acagctggtg gacttgaagc tgacagtgga
     3541 tgggctggag aaggaacgtg acttctactt cagcaaactt cgtgacatcg agctcatctg
     3601 ccaggagcat gaaagtgaaa acagccctgt tatctcaggc atcattggca tcctctatgc
     3661 cacagaggaa ggattcgcac cccctgagga cgatgagatt gaagagcatc aacaagaaga
     3721 ccaggacgag tacaccggtt catcgggatc catggtgagc gagctgatta aggagaacat
     3781 gcacatgaag ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga
     3841 gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggcgg tcgagggcgg
     3901 ccctctcccc ttcgccttcg acatcctggc taccagcttc atgtacggca gcaaaacctt
     3961 catcaaccac acccagggca tccccgactt ctttaagcag tccttccccg agggcttcac
     4021 atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag
     4081 cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa
     4141 cggccctgtg atgcagaaga aaacactcgg ctgggaggcc tccaccgaga ccctgtaccc
     4201 cgctgacggc ggcctggaag gcagagccga catggccctg aagctcgtgg gcgggggcca
     4261 cctgatctgc aacttgaaga ccacatacag atccaagaaa cccgctaaga acctcaagat
     4321 gcccggcgtc tactatgtgg acagaagact ggaaagaatc aaggaggccg acaaagagac
     4381 ctacgtcgag cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg
     4441 gcacagatga gacccagctt tcttgtacaa agttgatgta gatccagaca tgataagata
     4501 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     4561 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     4621 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt
     4681 cgcggccgcg gcgccaatgc attgggccct cgagcatatg agatctgtcc cgatctttct
     4741 cttctgtgct gtcaatttag agtagttttt gaggtaattt gggtgcattc tctgtaaaaa
     4801 caacaaataa atatttatat tgtatgcaat aaatgtaaca attacacata cactctctga
     4861 ttacagcatt aaagggttag ttcacccaaa aatgaaaaga tgtcattaat gactcaccct
     4921 catgtcgttc caagcccgta acacctccgt tcatcttcgg aacacagttt aagatatttt
     4981 agatttagtc cgagagcttt ctgtccctcc actgaaaatg tatgtacggt atactgtcca
     5041 tgtccagaaa ggtaataaaa acatcaaagt agtccatgtg acatcagtgg gttaggtaga
     5101 attatttgaa gcatcgaaaa tacattttgg tccaaaaata acaaaaccta cgactttatt
     5161 cagcattgta gcgtcactgc gaagtcgtga acgcggattg acaacagacc cggaagagaa
     5221 tacaatgccg aataaagtcg taggttttgt tatttttgga ccaaaatgta ttcgatgctt
     5281 caaaaaattc taactaaccc actgatgtca catggactac tttgatgttt ttattacctt
     5341 tctggacatg gacagtatac cgtacataca ttctcaatgg aggcacagaa agctctcgga
     5401 ctaaatctaa aatatcttaa actgtgttct gaagatgaac ggaggtctta cgggcttgga
     5461 acgacatgag ggcgagtcat taatgacatt attttcattt ttgggtgaac taacccttta
     5521 ataaaactga catttgcagc caattgcaca ctttataatg ttaaatactt gttgatttta
     5581 gaaatacagt agcatttagt gacaatcaaa cattaatttt atttctgatt atattgacaa
     5641 aaacacagga ctgtaaggct gacatgacag ggtgaattga tttcattaca ttagttttga
     5701 tggtaacact tttacaataa ggttcgttag ttaactacat tagttaacat gaactaataa
     5761 tgaactgcac gtatacatgt taatttcaac atttactaat actttatatc ttgttaacat
     5821 tagttaatgc actgtgaact aacatgaaca aacaatgaac atctgtattt ctattaacta
     5881 acgttaacaa agattaacag atacagtaac aaatgtattg ctcatggtta tttaatgtta
     5941 gttaatacat gaactaatca acctcattgt aaagtgttac cgttttgaca atacaacatg
     6001 tttggaaaaa aatttctcgc ttggttttct tgtgttttgc atgaattcaa acacaatgag
     6061 agcacggaat gataatgatt attcaaagct gcaataggcg ccaaatttcc cgtggaccta
     6121 acgttaccaa ttacaaatgc attttagcat tttattacgt gagccttata ttgtcaatcg
     6181 catgcattag tgaaaatagg cctactacac atataaaagg cgcgccatca acgtcaaaac
     6241 atgcacaccc aaaaaaaaaa aaaaaaaaaa aaaaaaaaac tatcagtaaa acaaaatact
     6301 taatgtactt acctcaatat gcttccttag gtttgatggc gaacttttga aggccgatat
     6361 ttctttatta agcgggagac agaggacaca tttcatcttg aatgaatctt tatttacacc
     6421 acttaaagaa aagaattcgc gaagatacgg ccacgggtgc tcttgatcct gtggctgatt
     6481 ttggactgtg ctgctcgcag ctgctgatga atcacatact tcctccattt tcttccactg
     6541 attgactgtt ataatttccc taatttccag gtcaaggtgc tgtgcattgt ggtaatagat
     6601 gtgacatgac gtcacttcca aaggaccaat gaacatgtct gaccaatttc atataatgtg
     6661 aaaacgattt tcataggcag aataaataac atttaaatta aactgggcat cagcgcaatt
     6721 caattggttt ggtaatagca agggaaaata gaatgaagtg atctccaaaa aataagtact
     6781 ttttgactgt aaataaaatt gtaaggagta aaaagtactt ttttttctaa aaaaatgtaa
     6841 ttaagtaaaa gtaaaagtat tgatttttaa ttgtactcaa gtaaagtaaa aatccccaaa
     6901 aataatactt aagtacagta atcaagtaaa attactcaag tactttacac ctctggttct
     6961 tgacccccta ccttcagcaa gcccagcaga tccactagtt ctagagcggc cgccaccgcg
     7021 gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     7081 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     7141 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     7201 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7261 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     7321 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7381 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7441 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7501 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7561 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7621 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7681 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7741 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7801 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7861 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7921 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7981 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     8041 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     8101 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     8161 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8221 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8281 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8341 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8401 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8461 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8521 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8581 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8641 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8701 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8761 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8821 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8881 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8941 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     9001 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     9061 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9121 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9181 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtg
//
