LOCUS       sequence-200756-        5659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  heat shock inducible transgenesis, cytosolic mKate2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5659)
  AUTHORS   Strzyz PJ, Lee HO, Sidhaye J, Weber IP, Leung LC, Norden C
  TITLE     Interkinetic nuclear migration is centrosome independent and ensures
            apical cell division to maintain tissue integrity.
  JOURNAL   Dev Cell. 2015 Jan 26;32(2):203-19. doi: 
            10.1016/j.devcel.2014.12.001. Epub 2015 Jan 15.
  PUBMED    25600237
REFERENCE   2  (bases 1 to 5659)
  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"; doi: 
            "10.1016/j.devcel.2014.12.001"; journalName: "Dev Cell"; date: 
            "2015-01-26- 26"; volume: "32"; issue: "2"; pages: "203-19"
COMMENT     Sequence Label: HS-mKate2
FEATURES             Location/Qualifiers
     source          1..5659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..22
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      1820..1829
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1826..2524
                     /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"
     polyA_signal    2660..2784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2697..2716)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2754..2773
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2791..3246)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2928..2949)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3140..3159
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3273..3377
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3379..3736
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3379..3570
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3402..3422)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3587..3722
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3771..4565
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3771..4565
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3825..3844)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4435..4454
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4753..4772)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4797..4844
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(5173..5659,1..102)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
       61 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      121 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
      181 cctgattctg tggataaccg tattaccgcc atgcattagt tattactagc gctaccggac
      241 tcagatctcg agctcaagct tcgaattctg cagcccgggg gatcctnnnt caggggtgtc
      301 gcttggttat ttccaaaaat caaattaatt ttattaaact attagaacga gcatgttttg
      361 tctatatgct acagaagata aaaaataata ggagttaaca gttataaaac aacacacttt
      421 gtttctattg attgttgacc acactggggt ctcattaagt tagattaaag acacactaac
      481 tgggtcaaaa gcagcagatt gatttcatag caccagggta aactttctaa cacttttacg
      541 gcaatcatat acattaaaat taaatacaga ccacgactga acaaggagga tgatctccaa
      601 tattaaacaa agagacttgt gcctatttct ctgagggtaa acatgacctc tcaagttagc
      661 aagttgtttt taacactaca aaaatagtta agactgcaat cccagaataa agtattggtt
      721 ttaaccaatc aatatagtac agtaaacatc catttgtttt gttgaaacgt taaacaaatc
      781 tgaccaaagc tattagctta tataaaacag gtttgccttc tatgtagctg aaaacaccac
      841 aggcccgatt ttgctactgt gtaaaacatt tcagcaagat ttttttattg catttttttt
      901 actgaatcnt caaacatttt atcattttag tttgttcatt cattgcaact ggaaaaacaa
      961 cacatcacac aaccgcacat atttcagcaa taagtacaat aaaacactca aataaaaaaa
     1021 acattttaaa tctctttgta tttttgaccg ctgtttcgcg taatttcacg gtaaaactct
     1081 ggaaatctcc actacattcc tctcagcggc tcctctcaat gacagctgaa gaagtgacgc
     1141 ggctgcctgc tgtgttttga ttggtcgaat tcactggagg cttccagaac agtgtagagt
     1201 ctgaacgggt gcgcgctctg ctgtatttaa agggcgaaag agagaccgca gagaaactca
     1261 accgaagaga agcgacttga caaagaagaa aagagcagcc tgacaggact ttttccccga
     1321 cgaggtgttt attcgctcta tttaagaatc tactgtaagg taagtctcaa tatattgtac
     1381 tctattggct aatcagaatt atatagagat tatatgtact taatgtcaaa aaatcaactt
     1441 tgtatatgta atctttttac atgtggactg cctatgttca tcttatttta ggtctactag
     1501 aaaattatat ttcccgtttt cacaataagg atttttaaaa aaagcaatga acagacgggc
     1561 atttacttta tgttgctgac attattttat atgagcataa taaccataaa tactagcaaa
     1621 tgtcctaaat gaatttgtgt taatgttgtc tacaaaagaa aattagcgtt ttacttgtac
     1681 aactaataat aacttggtta ttaagagaat ttcacttgtt gactagaaaa atcctttcat
     1741 aatgaaacaa ttgcaccata aattgtataa atataaaatt aattctaatt gttttttttt
     1801 nnncctgcag tcgccggtcg ccaccatggt gagcgagctg attaaggaga acatgcacat
     1861 gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga
     1921 aggcaagccc tacgagggca cccagaccat gagaatcaag gcggtcgagg gcggccctct
     1981 ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcaaaa ccttcatcaa
     2041 ccacacccag ggcatccccg acttctttaa gcagtccttc cccgagggct tcacatggga
     2101 gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca
     2161 ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcccat ccaacggccc
     2221 tgtgatgcag aagaaaacac tcggctggga ggcctccacc gagaccctgt accccgctga
     2281 cggcggcctg gaaggcagag ccgacatggc cctgaagctc gtgggcgggg gccacctgat
     2341 ctgcaacttg aagaccacat acagatccaa gaaacccgct aagaacctca agatgcccgg
     2401 cgtctactat gtggacagaa gactggaaag aatcaaggag gccgacaaag agacctacgt
     2461 cgagcagcac gaggtggctg tggccagata ctgcgacctc cctagcaaac tggggcacag
     2521 atgagcggcc cgacccgggc ggccgcgact ctagatcata atcagccata ccacatttgt
     2581 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     2641 gaatgcaatt gttgttgtta acttgnttat tgcagcttat aatggttaca aataaagcaa
     2701 tagcatcaca aatttcnnna caaataaagc atttttttca ctgcattcta gttgtggttt
     2761 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     2821 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2881 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     2941 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3001 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3061 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3121 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     3181 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3241 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3301 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3361 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3421 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3481 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3541 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3601 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3661 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3721 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3781 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3841 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3901 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     3961 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4021 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4081 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4141 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4201 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4261 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     4321 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4381 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4441 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4501 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4561 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4621 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4681 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     4741 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     4801 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     4861 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4921 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4981 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     5041 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5101 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5161 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5221 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5281 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     5341 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5401 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5461 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5521 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5581 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5641 gcgcacgagg gagcttcca
//
