LOCUS       sequence-225113-        4000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  to test Cas9 gRNA efficiency or HR-capacity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4000)
  AUTHORS   Liberante FG, Lappin K, Barros EM, Vohhodina J, Grebien F, Savage 
            KI, Mills KI
  TITLE     Altered splicing and cytoplasmic levels of tRNA synthetases in 
            SF3B1-mutant myelodysplastic syndromes as a therapeutic 
            vulnerability.
  JOURNAL   Sci Rep. 2019 Feb 25;9(1):2678. doi: 10.1038/s41598-019-39591-7.
  PUBMED    30804405
REFERENCE   2  (bases 1 to 4000)
  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: "2019-02-25"; volume: "9(1):2678. doi"; pages: " 
            10.1038/s41598-019-39591-7"
COMMENT     Sequence Label: pmKaxxte
FEATURES             Location/Qualifiers
     source          1..4000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          813..945
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      975..984
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2173..2192)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    2202..2283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2227..2246
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(2481..3069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2561..2580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3150..3944)
                     /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             complement(3150..3944)
                     /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(3261..3280)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3871..3890
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagg
      721 tcgtttagtg aaccgtcaga tcactagtag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctc gactgatcac aggtaagtat caaggttaca agacaggttt
      841 aaggaggcca atagaaactg ggcttgtcga gacagagaag attcttgcgt ttctgatagg
      901 cacctattgg tcttactgac atccactttg cctttctctc cacaggggta ccgaagccgc
      961 tagcgctacc ggtcgccacc atggtgagcg agctgattaa ggagaacatg cacatgaagc
     1021 tgtacatgga gggcaccgtg aacaaccacc acttcaagtg cacatccgag ggcgaaggca
     1081 agccctacga gggcacccag accatgagaa tcaaggtcgt cgagggcggc cctctcccct
     1141 tcgccttcga catcctggct accagcttca tgtacggcag caaaaccttc atcaaccacc
     1201 ctcagggcat ccccgacttc tttaagcagt ccttccctga gggcttcaca tgggagagag
     1261 tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg
     1321 gctgcctcat ctacaacgtc aagattagag gggtgaactt cccagccaac ggccctgtga
     1381 tgcagaagaa aacactcggc tgggaggcct ccaccgagac gctgtacccc gctgacggcg
     1441 gcctggaagg cgcatgtgac atggccctga agctcgtggg cgggggccac ctgatctgca
     1501 acttggagac cacatacaga tccaagaaac ccgctaagaa cctcaagatg cccgggatcc
     1561 gaattcgata tcaagcttag atctagaatc aaggtcgtcg agggcggccc tctccccttc
     1621 gccttcgaca tcctggctac cagcttcatg tacggcagca aaaccttcat caaccaccct
     1681 cagggcatcc ccgacttctt taagcagtcc ttccctgagg gcttcacatg ggagagagtc
     1741 accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc
     1801 tgcctcatct acaacgtcaa gattagaggg gtgaacttcc cagccaacgg ccctgtgatg
     1861 cagaagaaaa cactcggctg ggaggcctcc accgagacgc tgtaccccgc tgacggcggc
     1921 ctggaaggcg catgtgacat ggccctgaag ctcgtgggcg ggggccacct gatctgcaac
     1981 ttggagacca catacagatc caagaaaccc gctaagaacc tcaagatgcc cggcgtctac
     2041 aacgtggaca ggagactgga aagaatcaag gaggccgaca atgagaccta cgtcgagcag
     2101 cacgaggtgg ctgtggccag atactctact ggtggcgctg gtgatggagg taaatgaggt
     2161 cgactcgagc tcgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2221 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2281 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     2341 ttttaaagca agtaaaacct ctacaaatgt ggtacttaag agggggagac caaagggcga
     2401 gacgttaagg cctcacgtga catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2461 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2521 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2581 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttacggg atacctgtcc
     2641 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2701 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2761 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2821 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2881 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2941 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3001 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3061 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3121 tcacgttaag ggattttggt catgccgtct cagaagaact cgtcaagaag gcgatagaag
     3181 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
     3241 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
     3301 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
     3361 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgctcgcc
     3421 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
     3481 tgatcgacaa gaccggcttc catccgagta cgtgctctct cgatgcgatg tttcgcttgg
     3541 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
     3601 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
     3661 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagtacagc tgcgcaagga
     3721 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttgcagttc attcagggca
     3781 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     3841 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     3901 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcataatatt attgaagcat
     3961 ttatcagggt tcgtctcgtc ccggtctcct cccatgcatg
//
