LOCUS       sequence-203783-        8346 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 8346)
  AUTHORS   Taverna E, Mora-Bermudez F, Strzyz PJ, Florio M, Icha J, Haffner C, 
            Norden C, Wilsch-Brauninger M, Huttner WB
  TITLE     Non-canonical features of the Golgi apparatus in bipolar epithelial 
            neural stem cells.
  JOURNAL   Sci Rep. 2016 Feb 16;6:21206. doi: 10.1038/srep21206.
  PUBMED    26879757
REFERENCE   2  (bases 1 to 8346)
  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: "Sci Rep."; 
            date: "2016-02-16"; volume: "6:21206. doi"; pages: " 
            10.1038/srep21206"
COMMENT     Sequence Label: hsp70: mKate2-DN Kif5a
FEATURES             Location/Qualifiers
     source          1..8346
                     /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(5243..5263)
                     /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(5271..5287)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    5303..5437
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5327..5346)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5381..5400
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(5876..6063,6064..6075)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(6109..6125)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6109..6125)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(6162..6182)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6162..6180)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6201..6217)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6214..6236)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6225..6241
                     /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(6249..6255,6256..6273,6274..6279))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6294..6315
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6432..6449)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6603..7191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6683..6702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7362..8153,8154..8222))
                     /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     7985..8004
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8223..8327)
                     /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 ggcttcctct atcgtggtgc gcatctcaga ggaagagcgg
     3421 cagaaatacg aggaagagat ccgcaaactc taccgccagc tggacgacaa ggatgatgag
     3481 atcaatctcc agtgtcagct ggtggagaag ctgaaggagc agattctgga tcaggatgag
     3541 ctgctggcct ccacccgcgg cgacagtgat aaggtacaga cggagctggg tcgtctgcag
     3601 acggagagcg agagcgccaa ggcggaggtg cgggaggtgc tgcaggcgct ggaggagctg
     3661 gccgtcaact acgaccagaa gagccaggag gtggaggaga agagtctgca caacaaacag
     3721 ctggccgagc agctcagcca caagatggcc aacctgatgg agctggaggc ggagctagcg
     3781 cagatgcagg aagtgagctc acagcagagg aagcgcatcg ctgatgtgtt gaacggcctg
     3841 atgaaggacc tcagcgagtt cagcagcatc gtcggaaacg gagaactcaa gctaccggtg
     3901 gagatcagcg gtgcgattga ggaggagttc accgtggcac gtctctacat cagcaagatt
     3961 aaatctgagg tgaagtcgat ggtgaagcgc tgccggcagc tggagaacat gcagctggag
     4021 tgccatcgca agatggagga gaccagccgg gagctgtcgt catgccaact gctcatctca
     4081 cagcatgagg caaagatccg ctctctcaca gaatacatgc agaacgtgga gcagaagaag
     4141 agacagctgg aggacaacta tgatgctctc agtgaagaac tctacacact gcagaactct
     4201 gagagtcatg tggcagagct ggatggagga gagaaggcgg acagagaggc tgatgggaaa
     4261 aaggccgctg ttcgtccagc tctgcatgcg gagtctcgtc agacgcagct cagccgcctg
     4321 cgggatgaaa ttaacgagaa gcagagactc atcgacgagc tcacagataa gaaccaggct
     4381 ctggagttgg agctggctca cgtgcgctcc agtttcagca atctgaagct gcaggacgac
     4441 agcaagagcg cctgtctgga gcagatgtcg tttcaacagg agagacacga gcagtccaaa
     4501 caggatctca agggccttga ggagactgtt gctcgggaac tccagaccct ccacaacctg
     4561 cgcaagctgt tcgttcaaga cctcacgact cgggttaaaa aaagttcaga aattggacct
     4621 gatgatagtg gggggtctaa cacccagaag cagaagattt cctttcttga gaacaacctg
     4681 gatcagctta caaaggttca taaacagctg gtacgtgata atgcagatct ccgctgtgag
     4741 cttccgaagc tggagaaacg tcttcgttct acggctgagc gggttcgggc gctggagaca
     4801 gcactgaagg acgccaaaca gggcgcgatg aatgaccggc gccggtacca gcaggaggtg
     4861 gagcgcatcc gggacgccat gaggctgcgt tacccactgc gccgccccaa cgccgctcag
     4921 atcgctaagc cagtgagacc cgggcatcac gtcaccgcca catcgcccac cagcttctcc
     4981 gtcacacgcg caagcaaacc ggccacgtct tacagcaatg cccgattcct gcaggacgag
     5041 aagatggaga acagccagaa gaccaacact gttggataca gcactgtgag atccatagac
     5101 atactgagct cctgtccgct cgacgtggag aacggaaaca gcacagatat taatgataac
     5161 aggagtgatg ttcactgcgg cagcatggtg gacgatgcct ctcgagtgta catcatgcag
     5221 caggagactg cagccagtta agcaacttta ttatacatag ttgataattc actggccgtc
     5281 gttttacggt accatcgatg atgatccaga catgataaga tacattgatg agtttggaca
     5341 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     5401 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     5461 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     5521 atgtggtatg gctgattatg atcctctaga tcagatctgc gaagatacgg ccacgggtgc
     5581 tcttgatcct gtggctgatt ttggactgtg ctgctcgcag ctgctgatga atcacatact
     5641 tcctccattt tcttccactg attgactgtt ataatttccc taatttccag gtcaaggtgc
     5701 tgtgcattgt ggtaatagat gtgacatgac gtcacttcca aaggaccaat gaacatgtct
     5761 gaccaatttc atataatgtg aaaacgattt tcataggcag aataaataac atttaaatta
     5821 aactgggcat cagcgcaatt caattggttt ggtaatagca agggaaaata gaatgaagtg
     5881 atctccaaaa aataagtact ttttgactgt aaataaaatt gtaaggagta aaaagtactt
     5941 ttttttctaa aaaaatgtaa ttaagtaaaa gtaaaagtat tgatttttaa ttgtactcaa
     6001 gtaaagtaaa aatccccaaa aataatactt aagtacagta atcaagtaaa attactcaag
     6061 tactttacac ctctggttct tgacccccta ccttcagcaa gcccagcaga tccactagtt
     6121 ctagagcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt
     6181 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     6241 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     6301 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6361 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6421 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6481 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6541 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6601 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6661 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6721 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6781 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6841 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6901 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6961 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7021 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     7081 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7141 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7201 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta aggcattttg
     7261 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7321 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7381 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7441 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7501 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7561 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7621 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7681 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7741 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7801 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7861 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7921 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7981 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     8041 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     8101 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     8161 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     8221 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8281 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8341 aaagtg
//
