LOCUS       sequence-209372-        8144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Right TALEN for RBM20 with NN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8144)
  AUTHORS   Miyaoka Y, Chan AH, Judge LM, Yoo J, Huang M, Nguyen TD, Lizarraga 
            PP, So PL, Conklin BR
  TITLE     Isolation of single-base genome-edited human iPS cells without 
            antibiotic selection.
  JOURNAL   Nat Methods. 2014 Mar;11(3):291-3. doi: 10.1038/nmeth.2840. Epub 
            2014 Feb 9.
  PUBMED    24509632
REFERENCE   2  (bases 1 to 8144)
  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: 
            "10.1038/nmeth.2840"; journalName: "Nat Methods"; date: "2014-03"; 
            volume: "11"; issue: "3"; pages: "291-3"
COMMENT     Sequence Label: MR015-RBM20 Right TALEN
FEATURES             Location/Qualifiers
     source          1..8144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        384..587
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     538..558
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     632..651
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        632..650
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      670..679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             679..744
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             751..771
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2887..3474
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     primer_bind     complement(3504..3521)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3510..3734
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3780..4208
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3867..3886)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4077..4098
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4217..4237)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4222..4551
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4402..4537
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4464..4483
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4618..5412
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4672..4691)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5282..5301
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5586..5707
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5623..5642)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5677..5696
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5756..5772)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5756..5772)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5769..5791)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5780..5796
                     /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(5804..5810,5811..5828,5829..5834))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5849..5870
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5987..6004)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6158..6746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6238..6257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6917..7708,7709..7777))
                     /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     7540..7559
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7778..7882)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7957..7976)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
      601 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
      661 ctggctagcg ccaccatgga ctacaaagac catgacggtg attataaaga tcatgacatc
      721 gattacaagg atgacgatga caagatggcc cccaagaaga agaggaaggt gggcattcac
      781 cgcggggtac ctatggtgga cttgaggaca ctcggttatt cgcaacagca acaggagaaa
      841 atcaagccta aggtcaggag caccgtcgcg caacaccacg aggcgcttgt ggggcatggc
      901 ttcactcatg cgcatattgt cgcgctttca cagcaccctg cggcgcttgg gacggtggct
      961 gtcaaatacc aagatatgat tgcggccctg cccgaagcca cgcacgaggc aattgtaggg
     1021 gtcggtaaac agtggtcggg agcgcgagca cttgaggcgc tgctgactgt ggcgggtgag
     1081 cttagggggc ctccgctcca gctcgacacc gggcagctgc tgaagatcgc gaagagaggg
     1141 ggagtaacag cggtagaggc agtgcacgcc tggcgcaatg cgctcaccgg tgcccccctg
     1201 aacctgaccc cggaccaagt ggtggctatc gccagcaaca atggcggcaa gcaagcgctc
     1261 gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac cccggaccaa
     1321 gtggtggcta tcgccagcaa cattggcggc aagcaagcgc tcgaaacggt gcagcggctg
     1381 ttgccggtgc tgtgccagga ccatggcctg actccggacc aagtggtggc tatcgccagc
     1441 cacgatggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag
     1501 gaccatggcc tgactccgga ccaagtggtg gctatcgcca gccacgatgg cggcaagcaa
     1561 gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg
     1621 gaccaagtgg tggctatcgc cagcaacaat ggcggcaagc aagcgctcga aacggtgcag
     1681 cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt ggtggctatc
     1741 gccagcaaca atggcggcaa gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg
     1801 tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca cgatggcggc
     1861 aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga ccatggcctg
     1921 accccggacc aagtggtggc tatcgccagc aacggtggcg gcaagcaagc gctcgaaacg
     1981 gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgactccgga ccaagtggtg
     2041 gctatcgcca gccacgatgg cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg
     2101 gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc cagcaacatt
     2161 ggcggcaagc aagcgctcga aacggtgcag cggctgttgc cggtgctgtg ccaggaccat
     2221 ggcctgaccc cggaccaagt ggtggctatc gccagccacg atggcggcaa gcaagcgctc
     2281 gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac tccggaccaa
     2341 gtggtggcta tcgccagcca cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg
     2401 ttgccggtgc tgtgccagga ccatggcctg accccggacc aagtggtggc tatcgccagc
     2461 aacaatggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag
     2521 gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacaatgg cggcaagcaa
     2581 gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg
     2641 gaccaagtgg tggctatcgc cagcaacatt ggcggcaagc aagcgctcga aagcattgtg
     2701 gcccagctga gccggcctga tccggcgttg gccgcgttga ccaacgacca tctggtggcg
     2761 ttggcatgtc ttggtggacg acccgcgctc gatgcagtca aaaagggtct gcctcatgct
     2821 cccgcattga tcaaaagaac caaccggcgg attcccgaga gaacttccca tcgagtcgcg
     2881 ggatcccagc tggtgaagag cgagctggag gagaagaagt ccgagctgcg gcacaagctg
     2941 aagtacgtgc cccacgagta catcgagctg atcgagatcg ccaggaacag cacccaggac
     3001 cgcatcctgg agatgaaggt gatggagttc ttcatgaagg tgtacggcta caggggaaag
     3061 cacctgggcg gaagcagaaa gcctgacggc gccatctata cagtgggcag ccccatcgat
     3121 tacggcgtga tcgtggacac aaaggcctac agcggcggct acaatctgcc tatcggccag
     3181 gccgacgaga tgcagagata cgtggaggag aaccagaccc ggaataagca catcaacccc
     3241 aacgagtggt ggaaggtgta ccctagcagc gtgaccgagt tcaagttcct gttcgtgagc
     3301 ggccacttca agggcaacta caaggcccag ctgaccaggc tgaaccacat caccaactgc
     3361 aatggcgccg tgctgagcgt ggaggagctg ctgatcggcg gcgagatgat caaagccggc
     3421 accctgacac tggaggaggt gcggcgcaag ttcaacaacg gcgagatcaa cttctgataa
     3481 cttaagttta aaccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3541 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3601 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3661 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3721 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3781 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3841 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3901 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3961 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4021 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4081 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4141 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4201 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4261 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4321 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4381 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4441 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4501 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4561 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4621 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4681 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4741 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4801 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4861 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4921 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4981 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5041 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5101 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5161 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5221 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5281 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5341 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5401 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5461 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5521 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5581 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5641 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5701 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5761 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5821 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5881 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5941 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6001 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6061 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6121 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6181 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6241 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6301 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6361 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6421 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6481 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6541 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6601 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6661 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6721 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6781 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6841 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6901 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6961 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7021 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7081 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7141 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7201 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7261 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7321 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7381 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7441 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7501 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7561 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7621 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7681 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7741 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7801 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7861 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     7921 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     7981 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     8041 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     8101 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgc
//
