LOCUS       sequence-197978-        7943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALEN targeting upstream of HTT gene (KKR), forms obligate 
            heterodimer with USPQ-HTT-TALEN2.
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-TALEN1
FEATURES             Location/Qualifiers
     source          1..7943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      908..917
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             917..982
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             989..1009
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3951..3968)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3957..4181
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4227..4655
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4314..4333)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4524..4545
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4664..4684)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4669..4998
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4849..4984
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4911..4930
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5046..5093
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5112..5486
                     /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    5616..5737
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5653..5672)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5707..5726
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5786..5802)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5786..5802)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5799..5821)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5810..5826
                     /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(5834..5840,5841..5858,5859..5864))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5879..5900
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6017..6034)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6188..6776)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6268..6287)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6947..7738,7739..7807))
                     /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     7570..7589
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7808..7912)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagactcaa gctggctagt
      901 taagcttgcc accatggact acaaagacca tgacggtgat tataaagatc atgacatcga
      961 ttacaaggat gacgatgaca agatggcccc caagaagaag aggaaggtgg gcattcacgg
     1021 ggtacccgcc gctgtggatc tacgcacgct cggctacagc cagcagcaac aggagaagat
     1081 caaaccgaag gttcgttcga cagtggcgca gcaccacgag gcactggtcg gccatgggtt
     1141 tacacacgcg cacatcgttg cgctcagcca acacccggca gcgttaggga ccgtcgctgt
     1201 caagtatcag gacatgatcg cagcgttgcc agaggcgaca cacgaagcga tcgttggcgt
     1261 cggcaaacag tggtccggcg cacgcgctct ggaggccttg ctcacggtgg cgggagagtt
     1321 gagaggtcca ccgttacagt tggacacagg ccaacttctc aagattgcaa aacgtggcgg
     1381 cgtgaccgca gtggaggcag tgcatgcatg gcgcaatgca ctgacgggtg cccccctgaa
     1441 cttgacccca gaacaagtcg tcgcaatcgc gagcaataac ggcggaaaac aggctttgga
     1501 aacggtgcag aggctccttc cagtgctgtg tcaggcacat ggtctgactc cagaacaggt
     1561 tgtggccatc gcaagccacg acggaggaaa gcaagccctg gaaacagtac agaggctgtt
     1621 gcctgtcctg tgtcaagctc acggactcac tccagaacag gttgtggcca tcgcaagcca
     1681 cgacggagga aagcaagccc tggaaacagt acagaggctg ttgcctgtcc tgtgtcaagc
     1741 tcacggccta accccagaac aagtcgtcgc aatcgcgagc aataacggcg gaaaacaggc
     1801 tttggaaacg gtgcagaggc tccttccagt gctgtgtcag gcacatgggc ttaccccaga
     1861 acaagtcgtc gcaatcgcga gcaataacgg cggaaaacag gctttggaaa cggtgcagag
     1921 gctccttcca gtgctgtgtc aggcacatgg cttgacccca gaacaagtcg tcgcaatcgc
     1981 gagcaataac ggcggaaaac aggctttgga aacggtgcag aggctccttc cagtgctgtg
     2041 tcaggcacat ggactcactc cagaacaggt tgtggccatc gcaagccacg acggaggaaa
     2101 gcaagccctg gaaacagtac agaggctgtt gcctgtcctg tgtcaagctc acggcctaac
     2161 cccagaacaa gtcgtcgcaa tcgcgagcaa taacggcgga aaacaggctt tggaaacggt
     2221 gcagaggctc cttccagtgc tgtgtcaggc acatggtctg accccagaac aagtcgtcgc
     2281 aatcgcgagc aataacggcg gaaaacaggc tttggaaacg gtgcagaggc tccttccagt
     2341 gctgtgtcag gcacatggct tgaccccaga acaagtcgtc gcaatcgcga gcaataacgg
     2401 cggaaaacag gctttggaaa cggtgcagag gctccttcca gtgctgtgtc aggcacatgg
     2461 gcttacccct gagcaagtcg tcgcaatcgc ctccaacatt ggcgggaaac aggcactgga
     2521 gactgtccag cgcctgcttc ccgtgctttg ccaggcgcac ggcctaaccc cagaacaagt
     2581 cgtcgcaatc gcgagcaata acggcggaaa acaggctttg gaaacggtgc agaggctcct
     2641 tccagtgctg tgtcaggcac atggactcac ccctgagcaa gtcgtcgcaa tcgcctccaa
     2701 cattggcggg aaacaggcac tggagactgt ccagcgcctg cttcccgtgc tttgccaggc
     2761 gcacggcttg actccagaac aggttgtggc catcgcaagc cacgacggag gaaagcaagc
     2821 cctggaaaca gtacagaggc tgttgcctgt cctgtgtcaa gctcacgggc ttactccaga
     2881 acaggttgtg gccatcgcaa gccacgacgg aggaaagcaa gccctggaaa cagtacagag
     2941 gctgttgcct gtcctgtgtc aagctcacgg tctgacccca gaacaagtcg tcgcaatcgc
     3001 gagcaataac ggcggaaaac aggctttgga aacggtgcag aggctccttc cagtgctgtg
     3061 tcaggcacat ggcctaaccc cggagcaggt ggtggccatc gccagccacg acggcggcag
     3121 gccggcgctg gagagcattg ttgcccagtt atctcgccct gatccggcgt tggccgcgtt
     3181 gaccaacgac cacctcgtcg ccttggcctg cctcggcgga cgtcctgcgc tggatgcagt
     3241 gaaaaaggga ttgccgcacg cgccggcctt gatcaaaaga accaatcgcc gtattcccga
     3301 acgcacatcc catcgcgttg ccggatccca gctggtgaag tccgagctgg aggaaaagaa
     3361 gtccgaactg cgccacaagt tgaagtacgt ccctcacgag tacatcgaac tgatcgaaat
     3421 cgcccgcaac ccaacccagg accgtatcct ggagatgaag gtcatggagt tcttcatgaa
     3481 ggtgtacggt tatcgcggag agcatctcgg cggctctcgc aaacccgacg gcgctatcta
     3541 cacagtaggc tcccccatcg actacggcgt cattgtggac actaaggcct actcaggagg
     3601 ctacaacctt cccatcggcc aggccgacga aatgcagcgc tacgtgaagg agaatcagac
     3661 ccgcaacaag cacatcaacc ccaatgagtg gtggaaagtg tatccctcat ccgtcacaga
     3721 gttcaagttt ctgtttgtga gcggtcactt caagggtaac tacaaagccc agcttactcg
     3781 tctgaacagg aagaccaact gcaacggcgc cgtgctctca gtggaggagc tgctgatcgg
     3841 tggggagatg atcaaggccg ggaccctcac cttagaggag gtgcgccgga agttcaataa
     3901 tggagagatt aacttctgat aactcgagtc tagagaaacc cgctgatcag cctcgactgt
     3961 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     4021 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     4081 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     4141 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     4201 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     4261 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4321 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4381 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4441 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4501 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4561 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4621 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4681 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4741 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4801 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4861 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4921 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4981 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag
     5041 cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     5101 ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg
     5161 ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg
     5221 acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg
     5281 tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg
     5341 agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga
     5401 tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc
     5461 acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct
     5521 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5581 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5641 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5701 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5761 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5821 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5881 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5941 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6001 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6061 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6121 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6181 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6241 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6301 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6361 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6421 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6481 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6541 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6601 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6661 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6721 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6781 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6841 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6901 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6961 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7021 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7081 taccgcgaga gccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7141 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7201 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7261 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7321 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7381 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7441 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7501 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7561 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7621 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7681 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7741 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7801 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7861 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7921 cccgaaaagt gccacctgac gtc
//
