LOCUS       sequence-197512-        7943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALEN targeting upstream of HTT gene (ELD), forms obligate 
            heterodimer with USPQ-HTT-TALEN1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7943)
  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 7943)
  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: USPQ-HTT-TALEN2
FEATURES             Location/Qualifiers
     source          1..7943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..498
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        499..702
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     653..673
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     747..766
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        747..765
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      792..801
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             801..866
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             873..893
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3835..3852)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3841..4065
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4111..4539
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4198..4217)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4408..4429
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4548..4568)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4553..4882
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4733..4868
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4795..4814
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4930..4977
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4996..5370
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5500..5621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5537..5556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5591..5610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5670..5686)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5670..5686)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5683..5705)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5694..5710
                     /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(5718..5724,5725..5742,5743..5748))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5763..5784
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5901..5918)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6072..6660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6152..6171)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6831..7622,7623..7691))
                     /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     7454..7473
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7692..7796)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7871..7890)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
       61 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      121 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      181 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      241 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      301 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      361 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      421 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      481 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      541 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      601 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      661 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
      721 cccactgctt actggcttat cgaaattaat acgactcact atagggagac tcaagctggc
      781 tagttaagct tgccaccatg gactacaaag accatgacgg tgattataaa gatcatgaca
      841 tcgattacaa ggatgacgat gacaagatgg cccccaagaa gaagaggaag gtgggcattc
      901 acggggtacc cgccgctgtg gatctacgca cgctcggcta cagccagcag caacaggaga
      961 agatcaaacc gaaggttcgt tcgacagtgg cgcagcacca cgaggcactg gtcggccatg
     1021 ggtttacaca cgcgcacatc gttgcgctca gccaacaccc ggcagcgtta gggaccgtcg
     1081 ctgtcaagta tcaggacatg atcgcagcgt tgccagaggc gacacacgaa gcgatcgttg
     1141 gcgtcggcaa acagtggtcc ggcgcacgcg ctctggaggc cttgctcacg gtggcgggag
     1201 agttgagagg tccaccgtta cagttggaca caggccaact tctcaagatt gcaaaacgtg
     1261 gcggcgtgac cgcagtggag gcagtgcatg catggcgcaa tgcactgacg ggtgcccccc
     1321 tgaacttgac tccagaacag gttgtggcca tcgcaagcca cgacggagga aagcaagccc
     1381 tggaaacagt acagaggctg ttgcctgtcc tgtgtcaagc tcacggtctg accccagaac
     1441 aagtcgtcgc aatcgcgagc aataacggcg gaaaacaggc tttggaaacg gtgcagaggc
     1501 tccttccagt gctgtgtcag gcacatggac tcacccctga gcaagtcgtc gcaatcgcct
     1561 ccaacattgg cgggaaacag gcactggaga ctgtccagcg cctgcttccc gtgctttgcc
     1621 aggcgcacgg cctaacccct gagcaagtcg tcgcaatcgc ctccaacatt ggcgggaaac
     1681 aggcactgga gactgtccag cgcctgcttc ccgtgctttg ccaggcgcac gggcttaccc
     1741 cagaacaagt cgtcgcaatc gcgagcaata acggcggaaa acaggctttg gaaacggtgc
     1801 agaggctcct tccagtgctg tgtcaggcac atggcttgac cccagaacaa gtcgtcgcaa
     1861 tcgcgagcaa taacggcgga aaacaggctt tggaaacggt gcagaggctc cttccagtgc
     1921 tgtgtcaggc acatggactc actccagaac aggttgtggc catcgcaagc cacgacggag
     1981 gaaagcaagc cctggaaaca gtacagaggc tgttgcctgt cctgtgtcaa gctcacggcc
     2041 taactccaga acaggttgtg gccatcgcaa gccacgacgg aggaaagcaa gccctggaaa
     2101 cagtacagag gctgttgcct gtcctgtgtc aagctcacgg tctgacacct gagcaggttg
     2161 tggccattgc ctcgaatgga gggggcaaac aggcgttgga aaccgtacaa cgattgctgc
     2221 cggtcctttg ccaagcccac ggcttgacac ctgagcaggt tgtggccatt gcctcgaatg
     2281 gagggggcaa acaggcgttg gaaaccgtac aacgattgct gccggtcctt tgccaagccc
     2341 acgggcttac tccagaacag gttgtggcca tcgcaagcca cgacggagga aagcaagccc
     2401 tggaaacagt acagaggctg ttgcctgtcc tgtgtcaagc tcacggccta acccctgagc
     2461 aagtcgtcgc aatcgcctcc aacattggcg ggaaacaggc actggagact gtccagcgcc
     2521 tgcttcccgt gctttgccag gcgcacggac tcacacctga gcaggttgtg gccattgcct
     2581 cgaatggagg gggcaaacag gcgttggaaa ccgtacaacg attgctgccg gtcctttgcc
     2641 aagcccacgg cttgactcca gaacaggttg tggccatcgc aagccacgac ggaggaaagc
     2701 aagccctgga aacagtacag aggctgttgc ctgtcctgtg tcaagctcac gggcttaccc
     2761 ctgagcaagt cgtcgcaatc gcctccaaca ttggcgggaa acaggcactg gagactgtcc
     2821 agcgcctgct tcccgtgctt tgccaggcgc acggtctgac cccagaacaa gtcgtcgcaa
     2881 tcgcgagcaa taacggcgga aaacaggctt tggaaacggt gcagaggctc cttccagtgc
     2941 tgtgtcaggc acatggccta accccggagc aggtggtggc catcgccagc cacgacggcg
     3001 gcaggccggc gctggagagc attgttgccc agttatctcg ccctgatccg gcgttggccg
     3061 cgttgaccaa cgaccacctc gtcgccttgg cctgcctcgg cggacgtcct gcgctggatg
     3121 cagtgaaaaa gggattgccg cacgcgccgg ccttgatcaa aagaaccaat cgccgtattc
     3181 ccgaacgcac atcccatcgc gttgccggat cccagctggt gaagtccgag ctggaggaaa
     3241 agaagtccga actgcgccac aagttgaagt acgtccctca cgagtacatc gaactgatcg
     3301 aaatcgcccg caacccaacc caggaccgta tcctggagat gaaggtcatg gagttcttca
     3361 tgaaggtgta cggttatcgc ggagagcatc tcggcggctc tcgcaaaccc gacggcgcta
     3421 tctacacagt aggctccccc atcgactacg gcgtcattgt ggacactaag gcctactcag
     3481 gaggctacaa ccttcccatc ggccaggccg acgaaatgga gcgctacgtg gaggagaatc
     3541 agacccgcga caagcacctc aaccccaatg agtggtggaa agtgtatccc tcatccgtca
     3601 cagagttcaa gtttctgttt gtgagcggtc acttcaaggg taactacaaa gcccagctta
     3661 ctcgtctgaa ccacatcacc aactgcaacg gcgccgtgct ctcagtggag gagctgctga
     3721 tcggtgggga gatgatcaag gccgggaccc tcaccttaga ggaggtgcgc cggaagttca
     3781 ataatggaga gattaacttc tgataactcg agtctagaga aacccgctga tcagcctcga
     3841 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3901 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3961 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     4021 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     4081 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     4141 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4201 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4261 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4321 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4381 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4441 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4501 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     4561 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4621 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4681 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4741 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4801 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4861 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4921 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     4981 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
     5041 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
     5101 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
     5161 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
     5221 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
     5281 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
     5341 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
     5401 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5461 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     5521 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5581 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     5641 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5701 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5761 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5821 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5881 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5941 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6001 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6061 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6121 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6181 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6241 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6301 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6361 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6421 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6481 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6541 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6601 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6661 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6721 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6781 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6841 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6901 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6961 atgataccgc gagagccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7021 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7081 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7141 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7201 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7261 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7321 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7381 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7441 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7501 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7561 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7621 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7681 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7741 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7801 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     7861 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     7921 gtgtgttgga ggtcgctgag tag
//
