LOCUS       sequence-206120-        7452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALE vector with UNC5C-4 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 7452)
  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 7452)
  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: LLP251 pEF1a-UNC5C-4-SunTag
FEATURES             Location/Qualifiers
     source          1..7452
                     /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             3845..3865
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3908..3964
                     /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             3980..4036
                     /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             4052..4108
                     /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             4124..4180
                     /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             4196..4252
                     /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             4268..4324
                     /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             4340..4396
                     /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             4412..4468
                     /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             4484..4540
                     /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             4556..4612
                     /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          4667..4896
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     4879..4898
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     polyA_signal    5075..5196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5112..5131)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5298..5314)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5298..5314)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5311..5333)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5322..5338
                     /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(5346..5352,5353..5370,5371..5376))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5391..5412
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5529..5546)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5700..6288)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5780..5799)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6459..7250,7251..7319))
                     /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     7082..7101
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7320..7424)
                     /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 ccacgacgga ggcaaacagg ccctcgagac agtgcagcgg ctgctgcctg tgctgtgcca
     2221 ggcccacgga cttaccccag agcaggtcgt ggcaattgcc agcaatcacg ggggcaagca
     2281 ggccctcgaa acagtccaga gactgctgcc cgtgctgtgc caggcccacg gcctgacccc
     2341 agagcaggtc gtggcaattg ccagcaacgg cgggggaaag caggccctgg aaaccgtcca
     2401 gagactccta cctgtgctgt gccaggccca cggsctracc ccagagcagg tcgtggcaat
     2461 tgccagccac gacggaggca aacaggccct cgagacagtg cagcggctgc tgcctgtgct
     2521 gtgccaggcc cacgggctaa ccccagagca ggtcgtggca attgccagcc acgacggagg
     2581 caaacaggcc ctcgagacag tgcagcggct gctgcctgtg ctgtgccagg cccacggact
     2641 taccccagag caggtcgtgg caattgccag caacatcggc ggaaagcagg ctctggagac
     2701 agtgcagaga ctgctccccg tgctgtgcca ggcccacggc ctgaccccag agcaggtcgt
     2761 ggcaattgcc agcaacggcg ggggaaagca ggccctggaa accgtccaga gactcctacc
     2821 tgtgctgtgc caggcccacg ggctaacccc agagcaggtc gtggcaattg ccagcaacat
     2881 cggcggaaag caggctctgg agacagtgca gagactgctc cccgtgctgt gccaggccca
     2941 cggacttacc ccagagcagg tcgtggcaat tgccagccac gacggaggca aacaggccct
     3001 cgagacagtg cagcggctgc tgcctgtgct gtgccaggcc cacggcctga ccccagagca
     3061 ggtcgtggca attgccagca atcacggggg caagcaggcc ctcgaaacag tccagagact
     3121 gctgcccgtg ctgtgccagg cccacgggct aaccccagag caggtcgtgg caattgccag
     3181 caacggcggg ggaaagcagg ccctggaaac cgtccagaga ctcctacctg tgctgtgcca
     3241 ggcccacgga cttaccccag agcaggtcgt ggcaattgcc agcaatcacg ggggcaagca
     3301 ggccctcgaa acagtccaga gactgctgcc cgtgctgtgc caggcccacg gcctgacccc
     3361 agagcaggtc gtggcaattg ccagcaacgg cgggggaaag caggccctgg aaaccgtcca
     3421 gagactccta cctgtgctgt gccaggccca cgggctaacc ccagagcagg tcgtggcaat
     3481 tgccagcaat cacgggggca agcaggccct cgaaacagtc cagagactgc tgcccgtgct
     3541 gtgccaggcc cacggactta cgcctgagca ggtagtggct attgcatcca atcacggagg
     3601 cagacccgca ctggagtcaa tcgtggccca gctttcgagg ccggaccccg cgctggccgc
     3661 actcactaat gatcatcttg tagcgctggc ctgcctcggc ggacgacccg ccttggatgc
     3721 ggtgaagaag gggctcccgc acgcgcctgc attgattaag cggaccaaca gaaggattcc
     3781 cgagaggaca tcacatcgag tggcaggtgg cggagggtct gggggaggcg gtagtcaggc
     3841 cagcccgaaa aagaaaagaa aagtgggacg ggcgaaagga ggcggtggct caggcggtgg
     3901 gggaagcgaa gaacttttga gcaagaatta tcatcttgag aacgaagtgg ctcgtcttaa
     3961 gaaaggttct ggcagtggag aagaactgct ttcaaagaat taccacctgg aaaatgaggt
     4021 agctagactg aaaaagggga gcggaagtgg ggaggagttg ctgagcaaaa attatcattt
     4081 ggagaacgaa gtagcacgac taaagaaagg gtccggatcg ggtgaggagt tactctcgaa
     4141 aaattatcat ctcgaaaacg aagtggctcg gctaaaaaag ggcagtggtt ctggagaaga
     4201 gctattatct aaaaactacc acctcgaaaa tgaggtggca cgcttaaaaa agggaagtgg
     4261 cagtggtgaa gagctactat ccaagaatta tcatcttgag aacgaggtag cgcgtttgaa
     4321 gaagggttcc ggctcaggag aggaactgct ctcgaagaac tatcatcttg aaaatgaggt
     4381 cgctcgatta aaaaagggat cgggcagtgg tgaggaacta ctttcaaaga attaccacct
     4441 cgaaaacgaa gtagctcgat taaagaaagg ttcagggtcg ggtgaagaat tactgagtaa
     4501 aaattatcat ctggaaaatg aggtagcgag actaaaaaag gggagtggtt ctggcgagga
     4561 attgctatcg aaaaattatc atcttgagaa cgaagttgct aggctcaaaa agtgaggggg
     4621 aggctaactg gaattaattc gctgtctgcg agggccagct gttggggtga gtactccctc
     4681 tcaaaagcgg gcatgacttc tgcgctaaga ttgtcagttt ccaaaaacga ggaggatttg
     4741 atattcacct ggcccgcggt gatgcctttg agggtggccg cgtccatctg gtcagaaaag
     4801 acaatctttt tgttgtcaag cttgaggtgt ggcaggcttg agatctggcc atacacttga
     4861 gtgacaatga catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
     4921 aggtcgattt aaatgggaat tccacagcgg ccgcatgtct agataactga tcataatcag
     4981 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     5041 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     5101 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5161 tagttgtggt ttgtccaaac tcatcaatgt atcttaatgc gtcgagtgca ttctagttgt
     5221 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5281 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5341 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5401 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5461 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5521 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5581 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5641 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5701 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5761 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5821 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5881 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5941 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6001 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6061 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6121 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6181 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6241 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6301 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6361 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6421 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6481 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6541 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6601 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6661 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6721 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6781 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6841 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6901 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6961 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7021 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7081 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7141 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7201 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7261 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7321 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7381 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7441 gtgccacctg ac
//
