LOCUS       sequence-203779-        6657 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 6657)
  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 6657)
  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:mKate2-NWASP-CA
FEATURES             Location/Qualifiers
     source          1..6657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(514..525,526..688)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     1061..1077
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1097..1117
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    2637..2661
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2662..3357
                     /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(3359..3383)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3554..3574)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(3582..3598)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3614..3748
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3638..3657)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3692..3711
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(4187..4374,4375..4386)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4420..4436)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4420..4436)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4473..4493)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4473..4491)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4512..4528)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4525..4547)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4536..4552
                     /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(4560..4566,4567..4584,4585..4590))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4605..4626
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4743..4760)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4914..5502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4994..5013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5673..6464,6465..6533))
                     /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     6296..6315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6534..6638)
                     /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 tggcccacgc tggctaagaa
      241 ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg
      301 gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca
      361 atgctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa
      421 tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc
      481 acacaggtca gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt
      541 ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat
      601 ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt
      661 attaaggaag taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa
      721 tctcaaaaaa cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta
      781 tttaattatc aaactataag acagacaatc taatgccagt acacgctact caaagttgta
      841 aaacctcaga tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc
      901 tcttttgggg ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg cagccgatgc
      961 gggaagaggt gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga
     1021 ctccctggtg tctgaaacac aggccagatg ggccctcgag caggaaacag ctatgaccat
     1081 gattacgcca agctatcaac tttgtataga aaagttgctt caggggtgtc gcttggttat
     1141 ttccaaaaat caaattaatt ttattaaact attagaacga gcatgttttg tctatatgct
     1201 acagaagata aaaaataata ggagttaaca gttataaaac aacacacttt gtttctattg
     1261 attgttgacc acactggggt ctcattaagt tagattaaag acacactaac tgggtcaaaa
     1321 gcagcagatt gatttcatag caccagggta aactttctaa cacttttacg gcaatcatat
     1381 acattaaaat taaatacaga ccacgactga acaaggagga tgatctccaa tattaaacaa
     1441 agagacttgt gcctatttct ctgagggtaa acatgacctc tcaagttagc aagttggttt
     1501 taacactaca aaaatagtta agactgcaat cccagaataa agtattggtt ttaaccaatc
     1561 aatatagtac agtaaacatc catttgtttt gttgaaacgt taaacaaatc tgaccaaagc
     1621 tattagctta tataaaacag gtttgccttc tatgtagctg aaaacaccac aggcccgatt
     1681 ttgctactgt gtaaaacatt tcagcaagat ttttttattg catttttttt tactgaatcg
     1741 ttcaaacatt ttatcatttt agtttgttca ttcattgcaa ctggaaaaac aacacatcac
     1801 acaaccgcac atatttcagc aataagtaca ataaaacact caaataaaaa aaacatttta
     1861 aatctctttg tatttttgac cgctgtttcg cgtaatttca cggtaaaact ctggaaatct
     1921 ccactacatt cctctcagcg gctcctctca atgacagctg aagaagtgac gcggctgcct
     1981 gctgtgtttt gattggtcga attcactgga ggcttccaga acagtgtaga gtctgaacgg
     2041 gtgcgcgctc tgctgtattt aaagggcgaa agagagaccg cagagaaact caaccgaaga
     2101 gaagcgactt gacaaagaag aaaagagcag cctgacagga ctttttcccc gacgaggtgt
     2161 ttattcgctc tatttaagaa tctactgtaa ggtaagtctc aatatattgt actctattgg
     2221 ctaatcagaa ttatatagag attatatgta cttaatgtca aaaaatcaac tttgtatatg
     2281 taatcttttt acatgtggac tgcctatgtt catcttattt taggtctact agaaaattat
     2341 atttcccgtt ttcacaataa ggatttttaa aaaaagcaat gaacagacgg gcatttactt
     2401 tatgttgctg acattatttt atatgagcat aataaccata aatactagca aatgtcctaa
     2461 atgaatttgt gttaatgttg tctacaaaag aaaattagcg ttttacttgt acaactaata
     2521 ataacttggt tattaagaga atttcacttg ttgactagaa aaatcctttc ataatgaaac
     2581 aattgcacca taaattgtat aaatataaaa ttaattctaa ttgttttttt ttcctgccaa
     2641 gtttgtacaa aaaagcaggc tatggtgagc gagctgatta aggagaacat gcacatgaag
     2701 ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc
     2761 aagccctacg agggcaccca gaccatgaga atcaaggcgg tcgagggcgg ccctctcccc
     2821 ttcgccttcg acatcctggc taccagcttc atgtacggca gcaaaacctt catcaaccac
     2881 acccagggca tccccgactt ctttaagcag tccttccccg agggcttcac atgggagaga
     2941 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
     3001 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg
     3061 atgcagaaga aaacactcgg ctgggaggcc tccaccgaga ccctgtaccc cgctgacggc
     3121 ggcctggaag gcagagccga catggccctg aagctcgtgg gcgggggcca cctgatctgc
     3181 aacttgaaga ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc
     3241 tactatgtgg acagaagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag
     3301 cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg gcacagaaac
     3361 ccagctttct tgtacaaagt ggctgtgtct gaatccccgg accccaaccc tccagctcaa
     3421 gaggaatcct cagagggaat tgtgggagct ttgatgatgg tgatgcagaa aaggagtaaa
     3481 gttatccatt catcagaaga cgaagaggat gatgggggcg aagaagaaga agatgatgaa
     3541 tgggatgact aagcaacttt attatacata gttgataatt cactggccgt cgttttacgg
     3601 taccatcgat gatgatccag acatgataag atacattgat gagtttggac aaaccacaac
     3661 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     3721 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     3781 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     3841 ggctgattat gatcctctag atcagatctg cgaagatacg gccacgggtg ctcttgatcc
     3901 tgtggctgat tttggactgt gctgctcgca gctgctgatg aatcacatac ttcctccatt
     3961 ttcttccact gattgactgt tataatttcc ctaatttcca ggtcaaggtg ctgtgcattg
     4021 tggtaataga tgtgacatga cgtcacttcc aaaggaccaa tgaacatgtc tgaccaattt
     4081 catataatgt gaaaacgatt ttcataggca gaataaataa catttaaatt aaactgggca
     4141 tcagcgcaat tcaattggtt tggtaatagc aagggaaaat agaatgaagt gatctccaaa
     4201 aaataagtac tttttgactg taaataaaat tgtaaggagt aaaaagtact tttttttcta
     4261 aaaaaatgta attaagtaaa agtaaaagta ttgattttta attgtactca agtaaagtaa
     4321 aaatccccaa aaataatact taagtacagt aatcaagtaa aattactcaa gtactttaca
     4381 cctctggttc ttgaccccct accttcagca agcccagcag atccactagt tctagagcgg
     4441 ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     4501 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4561 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4621 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4681 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4741 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4801 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4861 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4921 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4981 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5041 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5101 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5161 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5221 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5281 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5341 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5401 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5461 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5521 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aaggcatttt ggtcatgaga
     5581 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5641 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5701 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5761 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5821 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5881 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5941 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6001 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6061 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6121 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6181 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6241 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6301 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6361 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6421 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6481 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6541 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6601 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtg
//
