LOCUS       sequence-177798-        7944 bp    DNA     circular SYN 15-APR-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 7944)
  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 7944)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..7944
                     /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..4999
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4850..4985
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4912..4931
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5047..5094
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5113..5487
                     /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    5617..5738
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5654..5673)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5708..5727
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5787..5803)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5787..5803)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5800..5822)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5811..5827
                     /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(5835..5841,5842..5859,5860..5865))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5880..5901
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6018..6035)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6189..6777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6269..6288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6948..7739,7740..7808))
                     /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     7571..7590
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7809..7913)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 ggagacccaa 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 cttgactcca gaacaggttg tggccatcgc aagccacgac ggaggaaagc aagccctgga
     1501 aacagtacag aggctgttgc ctgtcctgtg tcaagctcac ggtctgaccc cagaacaagt
     1561 cgtcgcaatc gcgagcaata acggcggaaa acaggctttg gaaacggtgc agaggctcct
     1621 tccagtgctg tgtcaggcac atggactcac ccctgagcaa gtcgtcgcaa tcgcctccaa
     1681 cattggcggg aaacaggcac tggagactgt ccagcgcctg cttcccgtgc tttgccaggc
     1741 gcacggccta acccctgagc aagtcgtcgc aatcgcctcc aacattggcg ggaaacaggc
     1801 actggagact gtccagcgcc tgcttcccgt gctttgccag gcgcacgggc 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 tccagaacag gttgtggcca tcgcaagcca cgacggagga aagcaagccc tggaaacagt
     2221 acagaggctg ttgcctgtcc tgtgtcaagc tcacggtctg acacctgagc aggttgtggc
     2281 cattgcctcg aatggagggg gcaaacaggc gttggaaacc gtacaacgat tgctgccggt
     2341 cctttgccaa gcccacggct tgacacctga gcaggttgtg gccattgcct cgaatggagg
     2401 gggcaaacag gcgttggaaa ccgtacaacg attgctgccg gtcctttgcc aagcccacgg
     2461 gcttactcca gaacaggttg tggccatcgc aagccacgac ggaggaaagc aagccctgga
     2521 aacagtacag aggctgttgc ctgtcctgtg tcaagctcac ggcctaaccc ctgagcaagt
     2581 cgtcgcaatc gcctccaaca ttggcgggaa acaggcactg gagactgtcc agcgcctgct
     2641 tcccgtgctt tgccaggcgc acggactcac acctgagcag gttgtggcca ttgcctcgaa
     2701 tggagggggc aaacaggcgt tggaaaccgt acaacgattg ctgccggtcc tttgccaagc
     2761 ccacggcttg actccagaac aggttgtggc catcgcaagc cacgacggag gaaagcaagc
     2821 cctggaaaca gtacagaggc tgttgcctgt cctgtgtcaa gctcacgggc ttacccctga
     2881 gcaagtcgtc gcaatcgcct ccaacattgg cgggaaacag gcactggaga ctgtccagcg
     2941 cctgcttccc gtgctttgcc aggcgcacgg 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 aatggagcgc tacgtggagg agaatcagac
     3661 ccgcgacaag cacctcaacc ccaatgagtg gtggaaagtg tatccctcat ccgtcacaga
     3721 gttcaagttt ctgtttgtga gcggtcactt caagggtaac tacaaagccc agcttactcg
     3781 tctgaaccac atcaccaact 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 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4441 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4501 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4561 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     4621 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4681 gggtgtggaa agtccccagg ctccccaggc aggcagaagt atgcaaagca tgcatctcaa
     4741 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4801 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4861 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4921 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     4981 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     5041 gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     5101 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
     5161 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
     5221 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
     5281 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
     5341 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
     5401 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
     5461 cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc
     5521 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5581 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     5641 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5701 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5761 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5821 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5881 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5941 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6001 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6061 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6121 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6181 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6241 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6301 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6361 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     6421 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6481 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6541 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6601 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     6661 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6721 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6781 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6841 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6901 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6961 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7021 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7081 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7141 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7201 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7261 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7321 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7381 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7441 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7501 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7561 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7621 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7681 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7741 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7801 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7861 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7921 ccccgaaaag tgccacctga cgtc
//
