LOCUS       sequence-418361-        5326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SB-transposable plasmid expressing a mutant histone H3.3-K27M in 
            tandem with Katushka (RFP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5326)
  TITLE     Castro lab plasmids Sep 2023
REFERENCE   2  (bases 1 to 5326)
  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"
COMMENT     Sequence Label: pKT-H3.3-K27M-katushka
FEATURES             Location/Qualifiers
     source          1..5326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1262..1834
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1285..1836
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1429..1446)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             1836..2540
                     /codon_start=1
                     /product="far-red-shifted derivative of red fluorescent 
                     protein from Entacmaea quadricolor, also known as Katushka 
                     (Shcherbo et al., 2007)"
                     /label=TurboFP635
                     /note="mammalian codon-optimized"
                     /translation="MVGEDSVLITENMHMKLYMEGTVNDHHFKCTSEGEGKPYEGTQTM
                     KIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERITTYEDGG
                     VLTATQDTSLQNGCLIYNVKINGVNFPSNGPVMQKKTLGWEASTEMLYPADSGLRGHSQ
                     MALKLVGGGYLHCSLKTTYRSKKPAKNLKMPGFYFVDRRLERIKEADKETYVEQHEMAV
                     ARYCDLPSKLGHS"
     polyA_signal    2619..2740
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2630..2649)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2684..2703
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2842..2860
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3170..3187)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3341..3929)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3421..3440)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4805..4824
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4883..4902
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(5243..5262)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5244..5262)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gagtcaatgg gaaaaaccca ttggagccaa gtacactgac tcaataggga ctttccattg
       61 ggttttgccc agtacataag gtcaataggg ggtgagtcaa caggaaagtc ccattggagc
      121 caagtacatt gagtcaatag ggactttcca atgggttttg cccagtacat aaggtcaatg
      181 ggaggtaagc caatgggttt ttcccattac tgacatgtat actgagtcat tagggacttt
      241 ccaatgggtt ttgcccagta cataaggtca ataggggtga atcaacagga aagtcccatt
      301 ggagccaagt acactgagtc aatagggact ttccattggg ttttgcccag tacaaaaggt
      361 caataggggg tgagtcaatg ggtttttccc attattggca catacataag gtcaataggg
      421 gtgactagtg gagaagagca tgcttgaggg ctgagtgccc ctcagtgggc agagagcaca
      481 tggcccacag tccctgagaa gttgggggga ggggtgggca attgaactgg tgcctagaga
      541 aggtggggct tgggtaaact gggaaagtga tgtggtgtac tggctccacc tttttcccca
      601 gggtggggga gaaccatata taagtgcagt agtctctgtg aacattcaag cttctgcctt
      661 ctccctcctg tgagtttggt aagtcactga ctgtctatgc ctgggaaagg gtgggcagga
      721 gatggggcag tgcaggaaaa gtggcactat gaaccctgca gccctaggaa tgcatctaga
      781 caattgtact aaccttcttc tctttcctct cctgacaggt tggtgtacag cggccgcatg
      841 gctcgtacaa agcagactgc ccgcaaatcg accggtggta aagcacccag gaagcaactg
      901 gctacaaaag ccgctcgcat gagtgcgccc tctactggag gggtgaagaa acctcatcgt
      961 tacaggcctg gtactgtggc gctccgtgaa attagacgtt atcagaagtc cactgaactt
     1021 ctgattcgca aacttccctt ccagcgtctg gtgcgagaaa ttgctcagga ctttaaaaca
     1081 gatctgcgct tccagagcgc agctatcggt gctttgcagg aggcaagtga ggcctatctg
     1141 gttggccttt ttgaagacac caacctgtgt gctatccatg ccaaacgtgt aacaattatg
     1201 ccaaaagaca tccagctagc acgccgcata cgtggagaac gtgcttaagg atccctcgag
     1261 gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     1321 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     1381 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     1441 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     1501 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     1561 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     1621 acgttgtgag ttggatagtt gtggaaagag tcaaatggct cacctcaagc gtattcaaca
     1681 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     1741 gcacatgctt tacatgtgtt tagtcgaggt taaaaaacgt ctaggccccc cgaaccacgg
     1801 ggacgtggtt ttcctttgaa aaacacgatg ataatatggt gggtgaggat agcgtgctga
     1861 tcaccgagaa catgcacatg aaactgtaca tggagggcac cgtgaacgac caccacttca
     1921 agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg aagatcaagg
     1981 tggtcgaggg cggccctctc cccttcgcct tcgacatcct ggctaccagc ttcatgtacg
     2041 gcagcaaaac ctttatcaac cacacccagg gcatccccga cttctttaag cagtccttcc
     2101 ctgagggctt cacatgggag aggatcacca catacgaaga cgggggcgtg ctgaccgcta
     2161 cccaggacac cagcctccag aacggctgcc tcatctacaa cgtcaagatc aacggggtga
     2221 acttcccatc caacggccct gtgatgcaga agaaaacact cggctgggag gccagcaccg
     2281 agatgctgta ccccgctgac agcggcctga gaggccatag ccagatggcc ctgaagctcg
     2341 tgggcggggg ctacctgcac tgctccctca agaccacata cagatccaag aaacccgcta
     2401 agaacctcaa gatgcccggc ttctacttcg tggacaggag actggaaaga atcaaggagg
     2461 ccgacaaaga gacctacgtc gagcagcacg agatggctgt ggccaggtac tgcgacctgc
     2521 ctagcaaact ggggcacagc tccggactca gataacgtac gtgcgcgcga atgcatccag
     2581 atctaggtac cagggcccag ctagctggcc agacatgata agatacattg atgagtttgg
     2641 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2701 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2761 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2821 caaatgtggt atggaattat caattaaccc tcactaaagg gagaccaagt taaacaattt
     2881 aaaggcaatg ctaccaaata ctaattgagt gtatgtaaac ttctgaccca ctgggaatgt
     2941 gatgaaagaa ataaaagctg aaatgaatca ttctctctac tattattctg atatttcaca
     3001 ttcttaaaat aaagtggtga tcctaactga cctaagacag ggaattttta ctaggattaa
     3061 atgtcaggaa ttgtgaaaaa gtgagtttaa atgtatttgg ctaaggtgta tgtaaacttc
     3121 cgacttcaac tgtataggga tctggtacca tttaaatctg ttccgcttcc tcgctcactg
     3181 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3241 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3301 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3361 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3421 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3481 cgcttaccgg atacctgtcc gcctttctcc cttcgggaac cgtggcgctt tctcatagct
     3541 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3601 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3661 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3721 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3781 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3841 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3901 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3961 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4021 tcttcaccta gatccttttt gccagtgtta caaccaatta accaattctg attagaaaaa
     4081 ctcatcgagc atcaaatgaa actgcaattt attccaattt attcatatca ggattatcaa
     4141 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     4201 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     4261 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     4321 ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc
     4381 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     4441 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     4501 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     4561 attcttctaa tacctggaat gctgtttttc cggggatcgc agtggtgagt aaccatgcat
     4621 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     4681 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     4741 acaactctgg cgcatcgggc ttcccataca agcgatagat tgtcgcacct gattgcccga
     4801 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     4861 gcctcgacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     4921 cagacagttt tattgttcat gatgcagctg gatccagatc cctatacagt tgaagtcgga
     4981 agtttacata cacttaagtt ggagtcatta aaactcgttt ttcaactact ccacaaattt
     5041 cttgttaaca aacaatagtt ttggcaagtc agttaggaca tctactttgt gcatgacaca
     5101 agtcattttt ccaacaattg tttacagaca gattatttca cttataattc actgtatcac
     5161 aattccagtg ggtcagaagt ttacatacac taagttgact gtgcctttaa acagcttgga
     5221 aaattccaga aaatgatggt ctccctatag tgagtcgtat tagatcccgg gattactagt
     5281 ttaaacctag gatgcatacc ggtcaagcta gcgataattc ctgcag
//
