LOCUS       sequence-206115-        6636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALE vector with DACH1-1 insert fused to SunTagx10, driven by EF1a, 
            with 3xHA tag, no PURO gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6636)
  AUTHORS   Pflueger C, Tan D, Swain T, Nguyen TV, Pflueger J, Nefzger C, Polo 
            JM, Ford E, Lister R
  TITLE     A modular dCas9-SunTag DNMT3A epigenome editing system overcomes 
            pervasive off-target activity of direct fusion dCas9-DNMT3A 
            constructs.
  JOURNAL   Genome Res. 2018 Jun 15. pii: gr.233049.117. doi: 
            10.1101/gr.233049.117.
  PUBMED    29907613
REFERENCE   2  (bases 1 to 6636)
  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"; journalName: "Genome Res.
            2018 Jun 15. pii: gr.233049.117. doi: 10.1101/gr.233049.117."
COMMENT     Sequence Label: LLP338 pEF1a-DACH1-1-SunTag
FEATURES             Location/Qualifiers
     source          1..6636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        284..1462
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          515..1453
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1410..1430
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1478..1504
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1508..1534
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1538..1564
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1574..1594
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3029..3049
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3092..3148
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             3164..3220
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             3236..3292
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             3308..3364
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             3380..3436
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             3452..3508
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             3524..3580
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             3596..3652
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             3668..3724
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             3740..3796
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     intron          3851..4080
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     4063..4082
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     polyA_signal    4259..4380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4296..4315)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4482..4498)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4482..4498)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4495..4517)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4506..4522
                     /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(4530..4536,4537..4554,4555..4560))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4575..4596
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4713..4730)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4884..5472)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4964..4983)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5643..6434,6435..6503))
                     /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     6266..6285
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6504..6608)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgtgcaaag atggataaag ttttaaacag agaggaatct
      241 ttgcagctaa tggaccttct aggtcttgaa aggagtggga attggctccg gtgcccgtca
      301 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
      361 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
      421 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
      481 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg
      541 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca
      601 gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg
      661 cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg
      721 ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc
      781 catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa
      841 tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc
      901 ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat
      961 cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg
     1021 tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag
     1081 atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga
     1141 gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc
     1201 atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg
     1261 gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga
     1321 gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc
     1381 cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt
     1441 tcttccattt caggtgtcgt gacgtacggc caccatgtac ccctatgacg tgcctgatta
     1501 cgccgcttat ccttacgatg tgcccgacta tgcagcctac ccatacgatg tgcccgacta
     1561 cgccgcaatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccgt
     1621 agatttgaga actttgggat attcacagca gcagcaggaa aagatcaagc ccaaagtgag
     1681 gtcgacagtc gcgcagcatc acgaagccct ggtgggtcat gggtttacac atgcccacat
     1741 cgtagccttg tcgcagcacc ctgcagccct tggcacggtc gccgtcaagt accaggacat
     1801 gattgcggcg ttgccggaag ccacacatga ggcgatcgtc ggtgtgggga aacagtggag
     1861 cggagcccgc gcccttgagg ccctgttgac ggtcgcggga gagctgagag ggcctcccct
     1921 tcagctggac acgggccagt tgctgaagat cgcgaagcgg ggaggagtca cggcggtcga
     1981 ggcggtgcac gcgtggcgca atgcgctcac gggagcaccc ctcaacctga ccccagagca
     2041 ggtcgtggca attgccagcc acgacggagg caaacaggcc ctcgagacag tgcagcggct
     2101 gctgcctgtg ctgtgccagg cccacgggct aaccccagag caggtcgtgg caattgccag
     2161 caacatcggc ggaaagcagg ctctggagac agtgcagaga ctgctccccg tgctgtgcca
     2221 ggcccacggm ctkaccccag agcaggtcgt ggcaattgcc agccacgacg gaggcaaaca
     2281 ggccctcgag acagtgcagc ggctgctgcc tgtgctgtgc caggcccacg grctwacccc
     2341 agagcaggtc gtggcaattg ccagcaaymw cggsggmaag caggcyctsg aracagtsca
     2401 gagactgcts cccgtgctgt gccaggccca cggmctkacc ccagagcagg tcgtggcaat
     2461 tgccagcaac ggcgggggaa agcaggccct ggaaaccgtc cagagactcc tacctgtgct
     2521 gtgccaggcc cacggsctra ccccagagca ggtcgtggca attgccagcc acgacggagg
     2581 caaacaggcc ctcgagacag tgcagcggct gctgcctgtg ctgtgccagg cccacgggct
     2641 aaccccagag caggtcgtgg caattgccag ccacgacgga ggcaaacagg ccctcgagac
     2701 agtgcagcgg ctgctgcctg tgctgtgcca ggcccacgga cttacgcctg agcaggtagt
     2761 ggctattgca tccaatcacg gaggcagacc cgcactggag tcaatcgtgg cccagctttc
     2821 gaggccggac cccgcgctgg ccgcactcac taatgatcat cttgtagcgc tggcctgcct
     2881 cggcggacga cccgccttgg atgcggtgaa gaaggggctc ccgcacgcgc ctgcattgat
     2941 taagcggacc aacagaagga ttcccgagag gacatcacat cgagtggcag gtggcggagg
     3001 gtctggggga ggcggtagtc aggccagccc gaaaaagaaa agaaaagtgg gacgggcgaa
     3061 aggaggcggt ggctcaggcg gtgggggaag cgaagaactt ttgagcaaga attatcatct
     3121 tgagaacgaa gtggctcgtc ttaagaaagg ttctggcagt ggagaagaac tgctttcaaa
     3181 gaattaccac ctggaaaatg aggtagctag actgaaaaag gggagcggaa gtggggagga
     3241 gttgctgagc aaaaattatc atttggagaa cgaagtagca cgactaaaga aagggtccgg
     3301 atcgggtgag gagttactct cgaaaaatta tcatctcgaa aacgaagtgg ctcggctaaa
     3361 aaagggcagt ggttctggag aagagctatt atctaaaaac taccacctcg aaaatgaggt
     3421 ggcacgctta aaaaagggaa gtggcagtgg tgaagagcta ctatccaaga attatcatct
     3481 tgagaacgag gtagcgcgtt tgaagaaggg ttccggctca ggagaggaac tgctctcgaa
     3541 gaactatcat cttgaaaatg aggtcgctcg attaaaaaag ggatcgggca gtggtgagga
     3601 actactttca aagaattacc acctcgaaaa cgaagtagct cgattaaaga aaggttcagg
     3661 gtcgggtgaa gaattactga gtaaaaatta tcatctggaa aatgaggtag cgagactaaa
     3721 aaaggggagt ggttctggcg aggaattgct atcgaaaaat tatcatcttg agaacgaagt
     3781 tgctaggctc aaaaagtgag ggggaggcta actggaatta attcgctgtc tgcgagggcc
     3841 agctgttggg gtgagtactc cctctcaaaa gcgggcatga cttctgcgct aagattgtca
     3901 gtttccaaaa acgaggagga tttgatattc acctggcccg cggtgatgcc tttgagggtg
     3961 gccgcgtcca tctggtcaga aaagacaatc tttttgttgt caagcttgag gtgtggcagg
     4021 cttgagatct ggccatacac ttgagtgaca atgacatcca ctttgccttt ctctccacag
     4081 gtgtccactc ccaggtccaa ctgcaggtcg atttaaatgg gaattccaca gcggccgcat
     4141 gtctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     4201 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     4261 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4321 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4381 atgcgtcgag tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4441 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4501 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4561 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4621 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4681 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4741 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4801 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4861 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4921 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4981 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5041 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5101 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5161 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5221 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5281 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5341 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5401 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5461 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5521 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5581 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5641 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5701 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5761 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5821 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5881 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5941 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6001 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6061 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6121 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6181 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6241 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6301 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6361 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6421 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6481 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6541 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6601 gttccgcgca catttccccg aaaagtgcca cctgac
//
