LOCUS       sequence-197977-        8742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALEN targeting downstream of HTT gene (KKR) with Blasticidin 
            selection, forms obligate heterodimer with DSPQ-HTT-TALEN2 BSD. 
            TALEN: HD, NI, NN, HD, NG, NG, HD, HD, NG, HD, NI, NN, HD, HD, NN, 
            HD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8742)
  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 8742)
  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-TALEN1 ZEO
FEATURES             Location/Qualifiers
     source          1..8742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1737..1802
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1809..1829
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    4799..5351
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4943..4960)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5170..5189
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5363..5761
                     /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(5765..5784)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5830..5885
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5884..5903)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6246..6262)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6246..6262)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6259..6281)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6270..6286
                     /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(6294..6300,6301..6318,6319..6324))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6339..6360
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6419..6614
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6465..6600
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6620..6754
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6657..6676)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6711..6730
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6822..6839)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6993..7581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7073..7092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7752..8543,8544..8612))
                     /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     8375..8394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8613..8717)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 ctrkrmsgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcagggact
     1741 acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat gacgatgaca
     1801 agatggcccc caagaagaag aggaaggtgg gcattcacgg ggtacccgcc gctgtggatc
     1861 tacgcacgct cggctacagc cagcagcaac aggagatgat caaaccgaag gttcgttcga
     1921 cagtggcgca gcaccacgag gcactggtcg gccatgggtt tacacacgcg cacatcgttg
     1981 cgctcagcca acacccggca gcgttaggga ccgtcgctgt caagtatcag gacatgatcg
     2041 cagcgttgcc agaggcgaca cacgaagcga tcgttggcgt cggcaaacag tggtccggcg
     2101 cacgcgctct ggaggccttg ctcacggtgg cgggagagtt gagaggtcca ccgttacagt
     2161 tggacacagg ccaacttctc aagattgcaa aacgtggcgg cgtgaccgca gtggaggcag
     2221 tgcatgcatg gcgcaatgca ctgacgggtg cccccctgaa cttgactcca gaacaggttg
     2281 tggccatcgc aagccacgac ggaggaaagc aagccctgga aacagtacag aggctgttgc
     2341 ctgtcctgtg tcaagctcac ggtctgaccc ctgagcaagt cgtcgcaatc gcctccaaca
     2401 ttggcgggaa acaggcactg gagactgtcc agcgcctgct tcccgtgctt tgccaggcgc
     2461 acggactcac cccagaacaa gtcgtcgcaa tcgcgagcaa taacggcgga aaacaggctt
     2521 tggaaacggt gcagaggctc cttccagtgc tgtgtcaggc acatggccta actccagaac
     2581 aggttgtggc catcgcaagc cacgacggag gaaagcaagc cctggaaaca gtacagaggc
     2641 tgttgcctgt cctgtgtcaa gctcacgggc ttacacctga gcaggttgtg gccattgcct
     2701 cgaatggagg gggcaaacag gcgttggaaa ccgtacaacg attgctgccg gtcctttgcc
     2761 aagcccacgg cttgacacct gagcaggttg tggccattgc ctcgaatgga gggggcaaac
     2821 aggcgttgga aaccgtacaa cgattgctgc cggtcctttg ccaagcccac ggactcactc
     2881 cagaacaggt tgtggccatc gcaagccacg acggaggaaa gcaagccctg gaaacagtac
     2941 agaggctgtt gcctgtcctg tgtcaagctc acggcctaac tccagaacag gttgtggcca
     3001 tcgcaagcca cgacggagga aagcaagccc tggaaacagt acagaggctg ttgcctgtcc
     3061 tgtgtcaagc tcacggtctg acacctgagc aggttgtggc cattgcctcg aatggagggg
     3121 gcaaacaggc gttggaaacc gtacaacgat tgctgccggt cctttgccaa gcccacggct
     3181 tgactccaga acaggttgtg gccatcgcaa gccacgacgg aggaaagcaa gccctggaaa
     3241 cagtacagag gctgttgcct gtcctgtgtc aagctcacgg gcttacccct gagcaagtcg
     3301 tcgcaatcgc ctccaacatt ggcgggaaac aggcactgga gactgtccag cgcctgcttc
     3361 ccgtgctttg ccaggcgcac ggcctaaccc cagaacaagt cgtcgcaatc gcgagcaata
     3421 acggcggaaa acaggctttg gaaacggtgc agaggctcct tccagtgctg tgtcaggcac
     3481 atggactcac tccagaacag gttgtggcca tcgcaagcca cgacggagga aagcaagccc
     3541 tggaaacagt acagaggctg ttgcctgtcc tgtgtcaagc tcacggcttg actccagaac
     3601 aggttgtggc catcgcaagc cacgacggag gaaagcaagc cctggaaaca gtacagaggc
     3661 tgttgcctgt cctgtgtcaa gctcacgggc ttaccccaga acaagtcgtc gcaatcgcga
     3721 gcaataacgg cggaaaacag gctttggaaa cggtgcagag gctccttcca gtgctgtgtc
     3781 aggcacatgg tctgactcca gaacaggttg tggccatcgc aagccacaac ggaggaaagc
     3841 aagccctgga aacagtacag aggctgttgc ctgtcctgtg tcaagctcac ggcctaaccc
     3901 cggagcaggt ggtggccatc gccagccacg acggcggcag gccggcgctg gagagcattg
     3961 ttgcccagtt atctcgccct gatccggcgt tggccgcgtt gaccaacgac cacctcgtcg
     4021 ccttggcctg cctcggcgga cgtcctgcgc tggatgcagt gaaaaaggga ttgccgcacg
     4081 cgccggcctt gatcaaaaga accaatcgcc gtattcccga acgcacatcc catcgcgttg
     4141 ccggatccca gctggtgaag tccgagctgg aggaaaagaa gtccgaactg cgccacaagt
     4201 tgaagtacgt ccctcacgag tacatcgaac tgatcgaaat cgcccgcaac ccaacccagg
     4261 accgtatcct ggagatgaag gtcatggagt tcttcatgaa ggtgtacggt tatcgcggag
     4321 agcatctcgg cggctctcgc aaacccgacg gcgctatcta cacagtaggc tcccccatcg
     4381 actacggcgt cattgtggac actaaggcct actcaggagg ctacaacctt cccatcggcc
     4441 aggccgacga aatgcagcgc tacgtgaagg agaatcagac ccgcaacaag cacatcaacc
     4501 ccaatgagtg gtggaaagtg tatccctcat ccgtcacaga gttcaagttt ctgtttgtga
     4561 gcggtcactt caagggtaac tacaaagccc agcttactcg tctgaacagg aagaccaact
     4621 gcaacggcgc cgtgctctca gtggaggagc tgctgatcgg tggggagatg atcaaggccg
     4681 ggaccctcac cttagaggag gtgcgccgga agttcaataa tggagagatt aacttctgat
     4741 aactcgagga tatcgcggcc gctacgtaaa ttccgccccc cccccccccc cccccctaac
     4801 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     4861 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     4921 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     4981 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     5041 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     5101 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     5161 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     5221 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     5281 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     5341 cgatgataat atggccacaa ccatggccaa gcctttgtct caagaagaat ccaccctcat
     5401 tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag
     5461 cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg
     5521 gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct
     5581 gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg
     5641 ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg
     5701 acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcta
     5761 aagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     5821 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     5881 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     5941 tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa
     6001 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     6061 tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc
     6121 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     6181 tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag cttggcgtaa
     6241 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6301 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6361 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc
     6421 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     6481 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     6541 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     6601 taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa
     6661 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6721 caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc
     6781 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6841 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6901 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6961 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7021 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7081 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7141 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7201 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7261 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7321 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7381 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     7441 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7501 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7561 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7621 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7681 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7741 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7801 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7861 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7921 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7981 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8041 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8101 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8161 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8221 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8281 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8341 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8401 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8461 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8521 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8581 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8641 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8701 caaatagggg ttccgcgcac atttccccga aaagtgccac ct
//
