LOCUS       sequence-203033-        6941 bp    DNA     circular SYN 14-APR-2025
DEFINITION  heat shock inducible transgenesis, stathmin stmn1 orig. from addgene
            #31326.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6941)
  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 6941)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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:Stathmin1-mKate2
FEATURES             Location/Qualifiers
     source          1..6941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     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(3112..3136)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3138..3836
                     /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(3838..3858)
                     /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(3866..3882)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3898..4032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3922..3941)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3976..3995
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4704..4720)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4704..4720)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4757..4777)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4757..4775)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4796..4812)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4809..4831)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4820..4836
                     /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(4844..4850,4851..4868,4869..4874))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4889..4910
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5027..5044)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5198..5786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5278..5297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5957..6748,6749..6817))
                     /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     6580..6599
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6818..6922)
                     /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 tggatggctt cttctgatat ccaggtgaaa gaactggaga
     2701 agcgtgcctc aggccaggct tttgagctga ttctcagccc tcggtcaaaa gaatctgttc
     2761 cagaattccc cctttcccct ccaaagaaga aggatctttc cctggaggaa attcagaaga
     2821 aattagaagc tgcagaagaa agacgcaagt cccatgaagc tgaggtcttg aagcagctgg
     2881 ctgagaaacg agagcacgag aaagaagtgc ttcagaaggc aatagaagag aacaacaact
     2941 tcagtaaaat ggcagaagag aaactgaccc acaaaatgga agctaataaa gagaaccgag
     3001 aggcacaaat ggctgccaaa ctggaacgtt tgcgagagaa ggataagcac attgaagaag
     3061 tgcggaagaa caaagaatcc aaagaccctg ctgacgagac tgaagctgac gacccagctt
     3121 tcttgtacaa agtgggaatg gtgagcgagc tgattaagga gaacatgcac atgaagctgt
     3181 acatggaggg caccgtgaac aaccaccact tcaagtgcac atccgagggc gaaggcaagc
     3241 cctacgaggg cacccagacc atgagaatca aggcggtcga gggcggccct ctccccttcg
     3301 ccttcgacat cctggctacc agcttcatgt acggcagcaa aaccttcatc aaccacaccc
     3361 agggcatccc cgacttcttt aagcagtcct tccccgaggg cttcacatgg gagagagtca
     3421 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct
     3481 gcctcatcta caacgtcaag atcagagggg tgaacttccc atccaacggc cctgtgatgc
     3541 agaagaaaac actcggctgg gaggcctcca ccgagaccct gtaccccgct gacggcggcc
     3601 tggaaggcag agccgacatg gccctgaagc tcgtgggcgg gggccacctg atctgcaact
     3661 tgaagaccac atacagatcc aagaaacccg ctaagaacct caagatgccc ggcgtctact
     3721 atgtggacag aagactggaa agaatcaagg aggccgacaa agagacctac gtcgagcagc
     3781 acgaggtggc tgtggccaga tactgcgacc tccctagcaa actggggcac agatgagcaa
     3841 ctttattata catagttgat aattcactgg ccgtcgtttt acggtaccat cgatgatgat
     3901 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3961 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     4021 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     4081 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcct
     4141 ctagatcaga tctgcgaaga tacggccacg ggtgctcttg atcctgtggc tgattttgga
     4201 ctgtgctgct cgcagctgct gatgaatcac atacttcctc cattttcttc cactgattga
     4261 ctgttataat ttccctaatt tccaggtcaa ggtgctgtgc attgtggtaa tagatgtgac
     4321 atgacgtcac ttccaaagga ccaatgaaca tgtctgacca atttcatata atgtgaaaac
     4381 gattttcata ggcagaataa ataacattta aattaaactg ggcatcagcg caattcaatt
     4441 ggtttggtaa tagcaaggga aaatagaatg aagtgatctc caaaaaataa gtactttttg
     4501 actgtaaata aaattgtaag gagtaaaaag tacttttttt tctaaaaaaa tgtaattaag
     4561 taaaagtaaa agtattgatt tttaattgta ctcaagtaaa gtaaaaatcc ccaaaaataa
     4621 tacttaagta cagtaatcaa gtaaaattac tcaagtactt tacacctctg gttcttgacc
     4681 ccctaccttc agcaagccca gcagatccac tagttctaga gcggccgcca ccgcggtgga
     4741 gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     4801 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4861 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4921 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4981 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5041 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5101 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5161 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5221 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5281 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5341 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5401 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5461 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5521 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5581 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5641 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5701 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5761 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5821 ctcagtggaa cgaaaactca cgttaaggca ttttggtcat gagattatca aaaaggatct
     5881 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5941 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6001 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6061 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6121 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6181 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6241 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6301 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6361 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6421 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6481 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6541 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6601 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6661 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6721 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6781 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6841 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6901 ataaacaaat aggggttccg cgcacatttc cccgaaaagt g
//
