LOCUS       sequence-197973-        8732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALEN targeting downstream of HTT gene (ELD) with Blasticidin 
            selection, forms obligate heterodimer with DSPQ-HTT-TALEN1 ZEO. 
            TALEN: NN, HD, NN, NN, HD, NG, NN, NI, NN, NN, HD, NI, NN, HD, NI, 
            NN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8732)
  AUTHORS   Ooi J, Langley SR, Xu X, Utami KH, Sim B, Huang Y, Harmston NP, Tay 
            YL, Ziaei A, Zeng R, Low D, Aminkeng F, Sobota RM, Ginhoux F, 
            Petretto E, Pouladi MA
  TITLE     Unbiased Profiling of Isogenic Huntington Disease hPSC-Derived CNS 
            and Peripheral Cells Reveals Strong Cell-Type Specificity of CAG 
            Length Effects.
  JOURNAL   Cell Rep. 2019 Feb 26;26(9):2494-2508.e7. doi: 
            10.1016/j.celrep.2019.02.008.
  PUBMED    30811996
REFERENCE   2  (bases 1 to 8732)
  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.celrep.2019.02"; journalName: "Cell Rep"; date: 
            "2019-02-26- 26"; volume: "26"; issue: "9"; pages: "2494-2508.e7"
COMMENT     Sequence Label: DSPQ-HTT-TALEN2 BSD
FEATURES             Location/Qualifiers
     source          1..8732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1728..1793
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1800..1820
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    join(4767..5341,5342..5344,5345..5353)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4934..4951)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5161..5180
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5354..5752
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(5756..5775)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5821..5876
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5875..5894)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6235..6251)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6235..6251)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6248..6270)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6259..6275
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6283..6289,6290..6307,6308..6313))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6328..6349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6408..6603
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6454..6589
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6609..6743
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6646..6665)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6700..6719
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6811..6828)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6982..7570)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7062..7081)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7741..8532,8533..8601))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8364..8383
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8602..8706)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcatggac tacaaagacc
     1741 atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac aagatggccc
     1801 ccaagaagaa gaggaaggtg ggcattcacg gggtacccgc cgctgtggat ctacgcacgc
     1861 tcggctacag ccagcagcaa caggagaaga tcaaaccgaa ggttcgttcg acagtggcgc
     1921 agcaccacga ggcactggtc ggccatgggt ttacacacgc gcacatcgtt gcgctcagcc
     1981 aacacccggc agcgttaggg accgtcgctg tcaagtatca ggacatgatc gcagcgttgc
     2041 cagaggcgac acacgaagcg atcgttggcg tcggcaaaca gtggtccggc gcacgcgctc
     2101 tggaggcctt gctcacggtg gcgggagagt tgagaggtcc accgttacag ttggacacag
     2161 gccaacttct caagattgca aaacgtggcg gcgtgaccgc agtggaggca gtgcatgcat
     2221 ggcgcaatgc actgacgggt gcccccctga acttgacccc agaacaagtc gtcgcaatcg
     2281 cgagcaataa cggcggaaaa caggctttgg aaacggtgca gaggctcctt ccagtgctgt
     2341 gtcaggcaca tggtctgact ccagaacagg ttgtggccat cgcaagccac gacggaggaa
     2401 agcaagccct ggaaacagta cagaggctgt tgcctgtcct gtgtcaagct cacggactca
     2461 ccccagaaca agtcgtcgca atcgcgagca ataacggcgg aaaacaggct ttggaaacgg
     2521 tgcagaggct ccttccagtg ctgtgtcagg cacatggcct aaccccagaa caagtcgtcg
     2581 caatcgcgag caataacggc ggaaaacagg ctttggaaac ggtgcagagg ctccttccag
     2641 tgctgtgtca ggcacatggg cttactccag aacaggttgt ggccatcgca agccacgacg
     2701 gaggaaagca agccctggaa acagtacaga ggctgttgcc tgtcctgtgt caagctcacg
     2761 gcttgacacc tgagcaggtt gtggccattg cctcgaatgg agggggcaaa caggcgttgg
     2821 aaaccgtaca acgattgctg ccggtccttt gccaagccca cggactcacc ccagaacaag
     2881 tcgtcgcaat cgcgagcaat aacggcggaa aacaggcttt ggaaacggtg cagaggctcc
     2941 ttccagtgct gtgtcaggca catggcctaa cccctgagca agtcgtcgca atcgcctcca
     3001 acattggcgg gaaacaggca ctggagactg tccagcgcct gcttcccgtg ctttgccagg
     3061 cgcacggtct gaccccagaa caagtcgtcg caatcgcgag caataacggc ggaaaacagg
     3121 ctttggaaac ggtgcagagg ctccttccag tgctgtgtca ggcacatggc ttgaccccag
     3181 aacaagtcgt cgcaatcgcg agcaataacg gcggaaaaca ggctttggaa acggtgcaga
     3241 ggctccttcc agtgctgtgt caggcacatg ggcttactcc agaacaggtt gtggccatcg
     3301 caagccacga cggaggaaag caagccctgg aaacagtaca gaggctgttg cctgtcctgt
     3361 gtcaagctca cggcctaacc cctgagcaag tcgtcgcaat cgcctccaac attggcggga
     3421 aacaggcact ggagactgtc cagcgcctgc ttcccgtgct ttgccaggcg cacggactca
     3481 ccccagaaca agtcgtcgca atcgcgagca ataacggcgg aaaacaggct ttggaaacgg
     3541 tgcagaggct ccttccagtg ctgtgtcagg cacatggctt gactccagaa caggttgtgg
     3601 ccatcgcaag ccacgacgga ggaaagcaag ccctggaaac agtacagagg ctgttgcctg
     3661 tcctgtgtca agctcacggg cttacccctg agcaagtcgt cgcaatcgcc tccaacattg
     3721 gcgggaaaca ggcactggag actgtccagc gcctgcttcc cgtgctttgc caggcgcacg
     3781 gtctgacccc agaacaagtc gtcgcaatcg cgagcaataa cggcggaaaa caggctttgg
     3841 aaacggtgca gaggctcctt ccagtgctgt gtcaggcaca tggcctaacc ccggagcagg
     3901 tggtggccat cgccagccac gacggcggca ggccggcgct ggagagcatt gttgcccagt
     3961 tatctcgccc tgatccggcg ttggccgcgt tgaccaacga ccacctcgtc gccttggcct
     4021 gcctcggcgg acgtcctgcg ctggatgcag tgaaaaaggg attgccgcac gcgccggcct
     4081 tgatcaaaag aaccaatcgc cgtattcccg aacgcacatc ccatcgcgtt gccggatccc
     4141 agctggtgaa gtccgagctg gaggaaaaga agtccgaact gcgccacaag ttgaagtacg
     4201 tccctcacga gtacatcgaa ctgatcgaaa tcgcccgcaa cccaacccag gaccgtatcc
     4261 tggagatgaa ggtcatggag ttcttcatga aggtgtacgg ttatcgcgga gagcatctcg
     4321 gcggctctcg caaacccgac ggcgctatct acacagtagg ctcccccatc gactacggcg
     4381 tcattgtgga cactaaggcc tactcaggag gctacaacct tcccatcggc caggccgacg
     4441 aaatggagcg ctacgtggag gagaatcaga cccgcgacaa gcacctcaac cccaatgagt
     4501 ggtggaaagt gtatccctca tccgtcacag agttcaagtt tctgtttgtg agcggtcact
     4561 tcaagggtaa ctacaaagcc cagcttactc gtctgaacca catcaccaac tgcaacggcg
     4621 ccgtgctctc agtggaggag ctgctgatcg gtggggagat gatcaaggcc gggaccctca
     4681 ccttagagga ggtgcgccgg aagttcaata atggagagat taacttctga taactcgagg
     4741 atatcgcggc cgctacgtaa attccgcccc tctccctccc ccccccctaa cgttactggc
     4801 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgtgattttc caccatattg
     4861 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     4921 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     4981 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     5041 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     5101 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     5161 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     5221 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     5281 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     5341 tatggccaca accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc
     5401 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct
     5461 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg
     5521 tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat
     5581 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt
     5641 gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac
     5701 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct aaagatcttt
     5761 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     5821 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     5881 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     5941 tggcaacata tgccatatgc tggctgccat gaacaaaggt ggctataaag aggtcatcag
     6001 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     6061 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     6121 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     6181 cctcttctct tatgaagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata
     6241 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6301 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6361 ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag
     6421 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     6481 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     6541 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     6601 aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6661 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6721 atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg
     6781 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6841 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6901 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6961 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7021 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7081 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7141 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     7201 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7261 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7321 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7381 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7441 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7501 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7561 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7621 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7681 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7741 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7801 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7861 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7921 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7981 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8041 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8101 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8161 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8221 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8281 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8341 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8401 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8461 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8521 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8581 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8641 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8701 tccgcgcaca tttccccgaa aagtgccacc tg
//
