LOCUS       Exported                5913 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Empty TALE vector fused to SunTagx10 driven by EF1a promoter, with 
            3xHA tag, no PURO gene.
ACCESSION   .
VERSION     .
KEYWORDS    LLP250 pEF1a-TALE-SunTag
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5913)
  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 5913)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5913
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             2306..2326
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2369..2425
                     /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             2441..2497
                     /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             2513..2569
                     /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             2585..2641
                     /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             2657..2713
                     /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             2729..2785
                     /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             2801..2857
                     /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             2873..2929
                     /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             2945..3001
                     /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             3017..3073
                     /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          3128..3357
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3340..3359
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     polyA_signal    3536..3657
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3573..3592)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3759..3775)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3759..3775)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3772..3794)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3783..3799
                     /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(3807..3837)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3852..3873
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3990..4007)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4161..4749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4241..4260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4920..5780)
                     /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     5543..5562
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5781..5885)
                     /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 cagagacgct
     2041 ggctgatggg cgtctcggag gcagacccgc actggagtca attgtggccc agctttcgag
     2101 gccggacccc gcgctggccg cactcactaa tgatcatctt gtagcgctgg cctgcctcgg
     2161 cggacgaccc gccttggatg cggtgaagaa ggggctcccg cacgcgcctg cattgattaa
     2221 gcggaccaac agaaggattc ccgagaggac atcacatcga gtggcaggtg gcggagggtc
     2281 tgggggaggc ggtagtcagg ccagcccgaa aaagaaaaga aaagtgggac gggcgaaagg
     2341 aggcggtggc tcaggcggtg ggggaagcga agaacttttg agcaagaatt atcatcttga
     2401 gaacgaagtg gctcgtctta agaaaggttc tggcagtgga gaagaactgc tttcaaagaa
     2461 ttaccacctg gaaaatgagg tagctagact gaaaaagggg agcggaagtg gggaggagtt
     2521 gctgagcaaa aattatcatt tggagaacga agtagcacga ctaaagaaag ggtccggatc
     2581 gggtgaggag ttactctcga aaaattatca tctcgaaaac gaagtggctc ggctaaaaaa
     2641 gggcagtggt tctggagaag agctattatc taaaaactac cacctcgaaa atgaggtggc
     2701 acgcttaaaa aagggaagtg gcagtggtga agagctacta tccaagaatt atcatcttga
     2761 gaacgaggta gcgcgtttga agaagggttc cggctcagga gaggaactgc tctcgaagaa
     2821 ctatcatctt gaaaatgagg tcgctcgatt aaaaaaggga tcgggcagtg gtgaggaact
     2881 actttcaaag aattaccacc tcgaaaacga agtagctcga ttaaagaaag gttcagggtc
     2941 gggtgaagaa ttactgagta aaaattatca tctggaaaat gaggtagcga gactaaaaaa
     3001 ggggagtggt tctggcgagg aattgctatc gaaaaattat catcttgaga acgaagttgc
     3061 taggctcaaa aagtgagggg gaggctaact ggaattaatt cgctgtctgc gagggccagc
     3121 tgttggggtg agtactccct ctcaaaagcg ggcatgactt ctgcgctaag attgtcagtt
     3181 tccaaaaacg aggaggattt gatattcacc tggcccgcgg tgatgccttt gagggtggcc
     3241 gcgtccatct ggtcagaaaa gacaatcttt ttgttgtcaa gcttgaggtg tggcaggctt
     3301 gagatctggc catacacttg agtgacaatg acatccactt tgcctttctc tccacaggtg
     3361 tccactccca ggtccaactg caggtcgatt taaatgggaa ttccacagcg gccgcatgtc
     3421 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3481 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3541 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3601 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaatg
     3661 cgtcgagtgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     3721 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     3781 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3841 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3901 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3961 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4021 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4081 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4141 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4201 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4261 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4321 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4381 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4441 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4501 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4561 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4621 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4681 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4741 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4801 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4861 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4921 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4981 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5041 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5101 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5161 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5221 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5281 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5341 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5401 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5461 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5521 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5581 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5641 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5701 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5761 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5821 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5881 ccgcgcacat ttccccgaaa agtgccacct gac
//
