LOCUS       sequence-209371-        8348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Left TALEN for RBM20 with NN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8348)
  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 8348)
  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 Left TALEN
FEATURES             Location/Qualifiers
     source          1..8348
                     /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             3091..3678
                     /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(3708..3725)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3714..3938
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3984..4412
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4071..4090)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4281..4302
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4421..4441)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4426..4755
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4606..4741
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4668..4687
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4822..5616
                     /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(4876..4895)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5486..5505
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5790..5911
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5827..5846)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5881..5900
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5960..5976)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5960..5976)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5973..5995)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5984..6000
                     /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(6008..6014,6015..6032,6033..6038))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6053..6074
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6191..6208)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6362..6950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6442..6461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7121..7912,7913..7981))
                     /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     7744..7763
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7982..8086)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8161..8180)
                     /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 gccagccacg 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 tgaccccgga ccaagtggtg gctatcgcca gcaacattgg cggcaagcaa
     1561 gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg
     1621 gaccaagtgg tggctatcgc cagcaacaat ggcggcaagc aagcgctcga aacggtgcag
     1681 cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt ggtggctatc
     1741 gccagcaaca ttggcggcaa gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg
     1801 tgccaggacc atggcctgac cccggaccaa gtggtggcta tcgccagcaa cggtggcggc
     1861 aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga ccatggcctg
     1921 accccggacc aagtggtggc tatcgccagc aacattggcg gcaagcaagc gctcgaaacg
     1981 gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga ccaagtggtg
     2041 gctatcgcca gcaacggtgg cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg
     2101 gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc cagcaacaat
     2161 ggcggcaagc aagcgctcga aacggtgcag cggctgttgc cggtgctgtg ccaggaccat
     2221 ggcctgaccc cggaccaagt ggtggctatc gccagcaaca atggcggcaa gcaagcgctc
     2281 gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac tccggaccaa
     2341 gtggtggcta tcgccagcca cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg
     2401 ttgccggtgc tgtgccagga ccatggcctg actccggacc aagtggtggc tatcgccagc
     2461 cacgatggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag
     2521 gaccatggcc tgactccgga ccaagtggtg gctatcgcca gccacgatgg cggcaagcaa
     2581 gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg
     2641 gaccaagtgg tggctatcgc cagcaacatt ggcggcaagc aagcgctcga aacggtgcag
     2701 cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt ggtggctatc
     2761 gccagcaaca atggcggcaa gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg
     2821 tgccaggacc atggcctgac cccggaccaa gtggtggcta tcgccagcaa cattggcggc
     2881 aagcaagcgc tcgaaagcat tgtggcccag ctgagccggc ctgatccggc gttggccgcg
     2941 ttgaccaacg accatctggt ggcgttggca tgtcttggtg gacgacccgc gctcgatgca
     3001 gtcaaaaagg gtctgcctca tgctcccgca ttgatcaaaa gaaccaaccg gcggattccc
     3061 gagagaactt cccatcgagt cgcgggatcc cagctggtga agagcgagct ggaggagaag
     3121 aagtccgagc tgcggcacaa gctgaagtac gtgccccacg agtacatcga gctgatcgag
     3181 atcgccagga acagcaccca ggaccgcatc ctggagatga aggtgatgga gttcttcatg
     3241 aaggtgtacg gctacagggg aaagcacctg ggcggaagca gaaagcctga cggcgccatc
     3301 tatacagtgg gcagccccat cgattacggc gtgatcgtgg acacaaaggc ctacagcggc
     3361 ggctacaatc tgcctatcgg ccaggccgac gagatgcaga gatacgtgga ggagaaccag
     3421 acccggaata agcacatcaa ccccaacgag tggtggaagg tgtaccctag cagcgtgacc
     3481 gagttcaagt tcctgttcgt gagcggccac ttcaagggca actacaaggc ccagctgacc
     3541 aggctgaacc acatcaccaa ctgcaatggc gccgtgctga gcgtggagga gctgctgatc
     3601 ggcggcgaga tgatcaaagc cggcaccctg acactggagg aggtgcggcg caagttcaac
     3661 aacggcgaga tcaacttctg ataacttaag tttaaaccgc tgatcagcct cgactgtgcc
     3721 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3781 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3841 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3901 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3961 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4021 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4081 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4141 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4201 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4261 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4321 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4381 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4441 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4501 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4561 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4621 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4681 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4741 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4801 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4861 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4921 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4981 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5041 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5101 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5161 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5221 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5281 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5341 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5401 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5461 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5521 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5581 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5641 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5701 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5761 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     5821 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5881 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5941 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6001 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6061 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6121 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6181 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6241 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6301 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6361 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6421 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6481 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6541 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6601 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6661 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6721 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6781 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6841 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6901 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6961 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7021 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7081 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7141 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7201 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7261 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7321 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7381 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7441 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7501 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7561 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7621 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7681 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7741 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7801 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7861 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7921 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7981 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8041 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8101 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     8161 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     8221 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     8281 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     8341 atatacgc
//
