LOCUS       sequence-203024-        5652 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 5652)
  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 5652)
  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..5652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..22
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      1816..1825
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1822..2520
                     /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    2656..2777
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2693..2712)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2747..2766
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2784..3239)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2921..2942)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3133..3152
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3266..3370
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3372..3729
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3372..3563
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3395..3415)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3580..3715
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3764..4558
                     /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             3764..4558
                     /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(3818..3837)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4428..4447
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4746..4765)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4790..4837
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(5166..5652,1..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 gatccttcag gggtgtcgct
      301 tggttatttc caaaaatcaa attaatttta ttaaactatt agaacgagca tgttttgtct
      361 atatgctaca gaagataaaa aataatagga gttaacagtt ataaaacaac acactttgtt
      421 tctattgatt gttgaccaca ctggggtctc attaagttag attaaagaca cactaactgg
      481 gtcaaaagca gcagattgat ttcatagcac cagggtaaac tttctaacac ttttacggca
      541 atcatataca ttaaaattaa atacagacca cgactgaaca aggaggatga tctccaatat
      601 taaacaaaga gacttgtgcc tatttctctg agggtaaaca tgacctctca agttagcaag
      661 ttgtttttaa cactacaaaa atagttaaga ctgcaatccc agaataaagt attggtttta
      721 accaatcaat atagtacagt aaacatccat ttgttttgtt gaaacgttaa acaaatctga
      781 ccaaagctat tagcttatat aaaacaggtt tgccttctat gtagctgaaa acaccacagg
      841 cccgattttg ctactgtgta aaacatttca gcaagatttt tttattgcat ttttttttac
      901 tgaatcgttc aaacatttta tcattttagt ttgttcattc attgcaactg gaaaaacaac
      961 acatcacaca accgcacata tttcagcaat aagtacaata aaacactcaa ataaaaaaaa
     1021 cattttaaat ctctttgtat ttttgaccgc tgtttcgcgt aatttcacgg taaaactctg
     1081 gaaatctcca ctacattcct ctcagcggct cctctcaatg acagctgaag aagtgacgcg
     1141 gctgcctgct gtgttttgat tggtcgaatt cactggaggc ttccagaaca gtgtagagtc
     1201 tgaacgggtg cgcgctctgc tgtatttaaa gggcgaaaga gagaccgcag agaaactcaa
     1261 ccgaagagaa gcgacttgac aaagaagaaa agagcagcct gacaggactt tttccccgac
     1321 gaggtgttta ttcgctctat ttaagaatct actgtaaggt aagtctcaat atattgtact
     1381 ctattggcta atcagaatta tatagagatt atatgtactt aatgtcaaaa aatcaacttt
     1441 gtatatgtaa tctttttaca tgtggactgc ctatgttcat cttattttag gtctactaga
     1501 aaattatatt tcccgttttc acaataagga tttttaaaaa aagcaatgaa cagacgggca
     1561 tttactttat gttgctgaca ttattttata tgagcataat aaccataaat actagcaaat
     1621 gtcctaaatg aatttgtgtt aatgttgtct acaaaagaaa attagcgttt tacttgtaca
     1681 actaataata acttggttat taagagaatt tcacttgttg actagaaaaa tcctttcata
     1741 atgaaacaat tgcaccataa attgtataaa tataaaatta attctaattg tttttttttc
     1801 ctgcagtcgc cggtcgccac catggtgagc gagctgatta aggagaacat gcacatgaag
     1861 ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc
     1921 aagccctacg agggcaccca gaccatgaga atcaaggcgg tcgagggcgg ccctctcccc
     1981 ttcgccttcg acatcctggc taccagcttc atgtacggca gcaaaacctt catcaaccac
     2041 acccagggca tccccgactt ctttaagcag tccttccccg agggcttcac atgggagaga
     2101 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
     2161 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg
     2221 atgcagaaga aaacactcgg ctgggaggcc tccaccgaga ccctgtaccc cgctgacggc
     2281 ggcctggaag gcagagccga catggccctg aagctcgtgg gcgggggcca cctgatctgc
     2341 aacttgaaga ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc
     2401 tactatgtgg acagaagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag
     2461 cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg gcacagatga
     2521 gcggcccgac ccgggcggcc gcgactctag atcataatca gccataccac atttgtagag
     2581 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     2641 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2701 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2761 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     2821 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     2881 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     2941 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3001 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3061 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3121 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3181 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3241 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3301 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3361 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3421 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3481 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3541 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3601 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3661 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3721 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3781 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3841 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3901 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     3961 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4021 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4081 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4141 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4201 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4261 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     4321 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4381 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4441 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4501 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4561 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4621 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4681 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     4741 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     4801 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     4861 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     4921 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     4981 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     5041 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5101 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5161 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5221 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5281 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5341 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5401 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5461 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5521 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5581 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5641 agggagcttc ca
//
