LOCUS       sequence-174236-        5732 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  HTT HR arms with 18 CAG repeats and piggyBac selection cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5732)
  AUTHORS   Xu X, Tay Y, Sim B, Yoon SI, Huang Y, Ooi J, Utami KH, Ziaei A, Ng 
            B, Radulescu C, Low D, Ng AY, Loh M, Venkatesh B, Ginhoux F, 
            Augustine GJ, Pouladi MA
  TITLE     Reversal of Phenotypic Abnormalities by CRISPR/Cas9-Mediated Gene 
            Correction in Huntington Disease Patient-Derived Induced Pluripotent
            Stem Cells.
  JOURNAL   Stem Cell Reports. 2017 Feb 21. pii: S2213-6711(17)30038-3. doi: 
            10.1016/j.stemcr.2017.01.022.
  PUBMED    28238795
REFERENCE   2  (bases 1 to 5732)
  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: "Stem Cell 
            Reports. 2017 Feb 21. pii: S2213-6711(17)30038-3. doi: 
            10.1016/j.stemcr.2017.01.022."
COMMENT     Sequence Label: pJOP-HTT-HR18Q
FEATURES             Location/Qualifiers
     source          1..5732
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     repeat_region   1005..1067
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1521..1732
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1745..2013
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      2042..2051
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2072..2734
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRC
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
     CDS             2804..2857
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2858..3454
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2858..2877)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3354..3374
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             3461..3514
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3515..4642
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="ASYPGHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRPEQKM
                     PTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQH
                     RLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDRH
                     PIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQRP
                     GERLDLAMLAAIRRVYGLLANTVRYLQCGGSWREDWGQLSGTAVPPQGAEPQSNAGPRP
                     HIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRSMHVFILDYDQSPAGCRDALL
                     QLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     polyA_signal    4691..4812
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4728..4747)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4782..4801
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   5569..5603
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 agccccgcgc cttcgcccgg gtggggcgct gcgctgtcag cggccttgct gtgtgaggca
       61 gaacctgcgg gggcaggggc gggctggttc cctggccagc cattggcaga gtccgcaggc
      121 tagggctgtc aatcatgctg gccggcgtgg ccccgcctcc gccggcgcgg ccccgcctcc
      181 gccggcgcag cgtctgggac gcaaggcgcc gtgggggctg ccgggacggg tccaagatgg
      241 acggccgctc aggttctgtt tttacctgcg gcccagagcc ccattcattg ccccggtgct
      301 gagcggcgcc gcgagtcggc ccgaggcctc cggggactgc cgtgccgggc gggagaccgc
      361 catggcgacc ctggaaaagc tgatgaaggc cttcgagtcc ctcaagtcct tccagcagca
      421 gcagcagcag cagcagcagc agcagcagca gcagcagcag caacagccgc caccgccgcc
      481 gccgccgccg ccgcctcctc acctaccaca cccgccgccg caggcacagc cgctgctgcc
      541 tcagccgcag ccgcccccgc cgccgccccc gccgccaccc ggcccggctg tggctgagga
      601 gccgctgcac cgaccgtgag tttgggcccg ctgcagctcc ctgtcccggc gggtcccagg
      661 ctacggcggg gatggcggta accctgcagc ctgcgggccg gcgacacgaa cccccggccc
      721 cgcagagaca gagtgaccca gcaacccaga gcccatgagg gacacccgcc ccctcctggg
      781 gcgaggcctt cccccacttc agccccgctc cctcacttgg gtcttccctt gtcctctcgc
      841 gaggggaggc agagccttgt tggggcctgt cctgaattca ccgaggggag tcacggcctc
      901 agccctctcg cccttcgcag gatgcgaaga gttggggcga gaacttgttt ctttttattt
      961 gcgagaaacc agggcggggg ttcttgcggc cgcggtctca ttaaccctag aaagataatc
     1021 atattgtgac gtacgttaaa gataatcatg cgtaaaattg acgcatgtgt tttatcggtc
     1081 tgtatatcga ggtttattta ttaatttgaa tagatattaa gttttattat atttacactt
     1141 acatactaat aataaattca acaaacaatt tatttatgtt tatttattta ttaaaaaaaa
     1201 acaaaaactc aaaatttctt ctataaagta acaaaacttt tatgagggac agcccccccc
     1261 caaagccccc agggatgtaa ttacgtccct cccccgctag ggggcagcag cgagccgccc
     1321 ggggctccgc tccggtccgg cgctcccccc gcatccccga gccggcagcg tgcggggaca
     1381 gcccgggcac ggggaaggtg gcacgggatc gctttcctct gaacgcttct cgctgctctt
     1441 tgagcctgca gacacctggg gggatacggg gaaaaggcct ccaaggccaa ggatctgcga
     1501 tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     1561 gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag
     1621 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     1681 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct
     1741 tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg
     1801 ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt
     1861 ctaggtaagt ttaaagctca ggtcgagaca gggcctttgt ccggcgctcc cttggagcct
     1921 acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt
     1981 tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc ctactctaga
     2041 cgccaccatg gagagcgacg agagcggcct gcccgccatg gagatcgagt gccgcatcac
     2101 cggcaccctg aacggcgtgg agttcgagct ggtgggcggc ggagagggca cccccaagca
     2161 gggccgcatg accaacaaga tgaaaagcac caaaggcgcc ctgaccttca gcccctacct
     2221 gctgagccac gtgatgggct acggcttcta ccacttcggc acctacccca gcggctacga
     2281 gaaccccttc ctgcacgcca tcaacaacgg cggctacacc aacacccgca tcgagaagta
     2341 cgaggacggc ggcgtgctgc acgtgagctt cagctaccgc tgcgaggccg gccgcgtgat
     2401 cggcgacttc aaggtggtgg gcaccggctt ccccgaggac agcgtgatct tcaccgacaa
     2461 gatcatccgc agcaacgcca ccgtggagca cctgcacccc atgggcgata acgtgctggt
     2521 gggcagcttc gcccgcacct tcagcctgcg cgacggcggc tactacagct tcgtggtgga
     2581 cagccacatg cacttcaaga gcgccatcca ccccagcatc ctgcagaacg ggggccccat
     2641 gttcgccttc cgccgcgtgg aggagctgca cagcaacacc gagctgggca tcgtggagta
     2701 ccagcacgcc ttcaagaccc ccatcgcctt cgccagatcc cgcgctcagt cgtccaattc
     2761 tgccgtggac ggcaccgccg gacccggctc caccggatct cgcgagggca gaggaagtct
     2821 tctaacatgc ggtgacgtgg aggagaatcc cggccctatg accgagtaca agcccacggt
     2881 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     2941 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     3001 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     3061 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     3121 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     3181 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     3241 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     3301 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acatccgcgc cccgcaacct
     3361 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     3421 gcgcacctgg tgcatgaccc gcaagcccgg tgccaccggt gagggcagag gaagtcttct
     3481 aacatgcggt gacgtggagg agaatcccgg ccctgcttcg taccccggcc atcaacacgc
     3541 gtctgcgttc gaccaggctg cgcgttctcg cggccatagc aaccgacgta cggcgttgcg
     3601 ccctcgccgg cagcaagaag ccacggaagt ccgcccggag cagaaaatgc ccacgctact
     3661 gcgggtttat atagacggtc cccacgggat ggggaaaacc accaccacgc aactgctggt
     3721 ggccctgggt tcgcgcgacg atatcgtcta cgtacccgag ccgatgactt actggcgggt
     3781 gctgggggct tccgagacaa tcgcgaacat ctacaccaca caacaccgcc tcgaccaggg
     3841 tgagatatcg gccggggacg cggcggtggt aatgacaagc gcccagataa caatgggcat
     3901 gccttatgcc gtgaccgacg ccgttctggc tcctcatatc gggggggagg ctgggagctc
     3961 acatgccccg cccccggccc tcaccctcat cttcgaccgc catcccatcg ccgccctcct
     4021 gtgctacccg gccgcgcggt accttatggg cagcatgacc ccccaggccg tgctggcgtt
     4081 cgtggccctc atcccgccga ccttgcccgg caccaacatc gtgcttgggg cccttccgga
     4141 ggacagacac atcgaccgcc tggccaaacg ccagcgcccc ggcgagcggc tggacctggc
     4201 tatgctggct gcgattcgcc gcgtttacgg gctacttgcc aatacggtgc ggtatctgca
     4261 gtgcggcggg tcgtggcggg aggactgggg acagctttcg gggacggccg tgccgcccca
     4321 gggtgccgag ccccagagca acgcgggccc acgaccccat atcggggaca cgttatttac
     4381 cctgtttcgg gcccccgagt tgctggcccc caacggcgac ctgtataacg tgtttgcctg
     4441 ggccttggac gtcttggcca aacgcctccg ttccatgcac gtctttatcc tggattacga
     4501 ccaatcgccc gccggctgcc gggacgccct gctgcaactt acctccggga tggtccagac
     4561 ccacgtcacc acccccggct ccataccgac gatatgcgac ctggcgcgca cgtttgcccg
     4621 ggagatgggg gaggctaact gattttcatc gatttctcga gcttctactg tctggttaga
     4681 ccgttaacta aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4741 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4801 caatgtatct tatcatgtct ggaattgact caaatgatgt caattagtct atcagaagct
     4861 catctccttc cgggggacaa gacatccctg tttaatattt aaacagcagt gttcccaaac
     4921 tgggttctta tatcccttgc tctggtcaac caggttgcag ggtttcctgt cctcacagga
     4981 acgaagtccc taaagaaaca gtggcagcca ggtttagccc cggaattgac tggattcctt
     5041 ttttagggcc cattggtatg gctttttccc cgtatccccc caggtgtctg caggctcaaa
     5101 gagcagcgag aagcgttcag aggaaagcga tcccgtgcca ccttccccgt gcccgggctg
     5161 tccccgcacg ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg
     5221 gcggctcgct gctgccccct agcgggggag ggacgtaatt acatccctgg gggctttggg
     5281 ggggggctgt ccctgatatc tataacaaga aaatatatat ataataagtt atcacgtaag
     5341 tagaacatga aataacaata taattatcgt atgagttaaa tcttaaaagt cacgtaaaag
     5401 ataatcatgc gtcattttga ctcacgcggt cgttatagtt caaaatcagt gacacttacc
     5461 gcattgacaa gcacgcctca cgggagctcc aagcggcgac tgagatgtcc taaatgcaca
     5521 gcgacggatt cgcgctattt agaaagagag agcaatattt caagaatgca tgcgtcaatt
     5581 ttacgcagac tatctttcta gggttaactg cgttgtgaag agaacttgga ggagccgaga
     5641 tttgctcagt gccacttccc tcttctagtc tgagagggaa gagggctggg ggcgcgggac
     5701 acttcgagag gaggcggggt ttggagctgg ag
//
