LOCUS       sequence-203776-        7776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  heat shock inducible transgenesis, primary cillium marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7776)
  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 7776)
  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:Arl13b-mKate2
FEATURES             Location/Qualifiers
     source          1..7776
                     /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"
     protein_bind    complement(3947..3971)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3973..4671
                     /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(4673..4693)
                     /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(4701..4717)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    4733..4867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4757..4776)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4811..4830
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(5306..5493,5494..5505)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(5539..5555)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5539..5555)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5592..5612)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5592..5610)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5631..5647)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5644..5666)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5655..5671
                     /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(5679..5685,5686..5703,5704..5709))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5724..5745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5862..5879)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6033..6621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6113..6132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6792..7583,7584..7652))
                     /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     7415..7434
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7653..7757)
                     /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 tggatgttcc agtctgatgg ccaactgctg caacttgttc
     2701 aagcggtggc gggagcctgt cagaaaggtg acacttgtga tggtcggcct tgataatgct
     2761 ggtaaaacag ccacagcaaa gggaatccaa ggagagcatc ctgaagacgt agctcccact
     2821 gttggctttt ctaaaattga tctgagacaa ggaaagttcc aagttaccat ctttgactta
     2881 ggaggtggaa aaagaattcg gggaatttgg aagaattatt atgctgaatc ctatggggta
     2941 atatttgttg tggattccag tgatgaggag agaatggaag aaacaaagga gacaatgtcc
     3001 gaagtgctaa gacacccgag gatatcagga aagcctatat tggtgctggc caacaagcaa
     3061 gacaaggagg gggctttagg cgaagcggat gtgattgagt gtctctcgct ggagaagctg
     3121 gtcaacgagc acaagtgcct gtgtcagatc gaaccttgtt cagcagtctt gggatatgga
     3181 aagaaaattg acaagtccat taaaaagggg ctttattggc tactgcatat cattgcaaag
     3241 gactttgatg ccttaagtga acgcatccag aaagacacaa ctgaacagcg agctcttgag
     3301 gaacaagaga aacgtgagag ggctgaacga gtccggaagt taagggaaga aagagaacga
     3361 gagcagacgg aacttgatgg gaccagtggt ctggctgaga ttgactcagg accagttctt
     3421 gcgaatcctt tccaacccat tgcagctgta atcattgaga atgaaaagaa gcaggaaaaa
     3481 gagaaaaaga agcagaccgt ggagaaggac agtgatgttg gtctcctaga acataaagtg
     3541 gagccagagc aagcagcgcc tcagagtgag gcagattgct gcctccaaaa ccctgacgag
     3601 cgagtggtgg acagttacag ggaggcactg tcgcagcagc tggacagtga ggatgagcag
     3661 gaccagcgag ggtcagagtc cggggaaaac agtaagaaga aaaccaagaa actaagaatg
     3721 aaaaggagtc atcgggtaga accagtcaat acagacgagt ctactccaaa gagtcccacg
     3781 cctccccaac ctccccctcc tgttggctgg ggaaccccca aagtcactag acttccaaaa
     3841 cttgagcctc ttggtgaaac acgtcacaat gatttctatg ggaagccgct gcctcccctg
     3901 gctgtgcgac agagacctaa cggtgatgct caggacacga tctcagaccc agctttcttg
     3961 tacaaagtgg gaatggtgag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg
     4021 gagggcaccg tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac
     4081 gagggcaccc agaccatgag aatcaaggcg gtcgagggcg gccctctccc cttcgccttc
     4141 gacatcctgg ctaccagctt catgtacggc agcaaaacct tcatcaacca cacccagggc
     4201 atccccgact tctttaagca gtccttcccc gagggcttca catgggagag agtcaccaca
     4261 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
     4321 atctacaacg tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag
     4381 aaaacactcg gctgggaggc ctccaccgag accctgtacc ccgctgacgg cggcctggaa
     4441 ggcagagccg acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttgaag
     4501 accacataca gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg
     4561 gacagaagac tggaaagaat caaggaggcc gacaaagaga cctacgtcga gcagcacgag
     4621 gtggctgtgg ccagatactg cgacctccct agcaaactgg ggcacagatg agcaacttta
     4681 ttatacatag ttgataattc actggccgtc gttttacggt accatcgatg atgatccaga
     4741 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     4801 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     4861 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     4921 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctaga
     4981 tcagatctgc gaagatacgg ccacgggtgc tcttgatcct gtggctgatt ttggactgtg
     5041 ctgctcgcag ctgctgatga atcacatact tcctccattt tcttccactg attgactgtt
     5101 ataatttccc taatttccag gtcaaggtgc tgtgcattgt ggtaatagat gtgacatgac
     5161 gtcacttcca aaggaccaat gaacatgtct gaccaatttc atataatgtg aaaacgattt
     5221 tcataggcag aataaataac atttaaatta aactgggcat cagcgcaatt caattggttt
     5281 ggtaatagca agggaaaata gaatgaagtg atctccaaaa aataagtact ttttgactgt
     5341 aaataaaatt gtaaggagta aaaagtactt ttttttctaa aaaaatgtaa ttaagtaaaa
     5401 gtaaaagtat tgatttttaa ttgtactcaa gtaaagtaaa aatccccaaa aataatactt
     5461 aagtacagta atcaagtaaa attactcaag tactttacac ctctggttct tgacccccta
     5521 ccttcagcaa gcccagcaga tccactagtt ctagagcggc cgccaccgcg gtggagctcc
     5581 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     5641 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5701 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5761 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5821 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5881 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5941 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6001 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6061 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6121 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6181 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6241 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6301 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6361 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6421 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6481 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6541 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6601 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6661 tggaacgaaa actcacgtta aggcattttg gtcatgagat tatcaaaaag gatcttcacc
     6721 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6781 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6841 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6901 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6961 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7021 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7081 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7141 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7201 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7261 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7321 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7381 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7441 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7501 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7561 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7621 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7681 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7741 caaatagggg ttccgcgcac atttccccga aaagtg
//
