LOCUS Exported 8121 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION TET1-catalytic domain (CD) expression vector for TALE-TET1 fusion protein to induce targeted demethylation in mammalian cells. ACCESSION . VERSION . KEYWORDS pJFA274 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8121) AUTHORS Maeder ML, Angstman JF, Richardson ME, Linder SJ, Cascio VM, Tsai SQ, Ho QH, Sander JD, Reyon D, Bernstein BE, Costello JF, Wilkinson MF, Joung JK TITLE Targeted DNA demethylation and activation of endogenous genes using programmable TALE-TET1 fusion proteins. JOURNAL Nat Biotechnol. 2013 Oct 9. doi: 10.1038/nbt.2726. PUBMED 24108092 REFERENCE 2 (bases 1 to 8121) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8121 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 331..350 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(569..673) /gene="bla" /label=AmpR promoter primer_bind complement(752..771) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 927..2105 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 1158..2096 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 2053..2073 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 2113..2122 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2125..2145 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(5161..5180) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5215..5234 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5271..5291) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5276..5605 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5456..5591 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5518..5537 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5615..6214 /codon_start=1 /gene="pac from Streptomyces" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA" primer_bind complement(5615..5634) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 6111..6131 /label=Puro-F /note="Puromycin resistance gene, forward primer" CDS 6276..6317 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 6327..6344 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(6367..6384) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6373..6597 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(6668..6684) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6668..6684) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6681..6703) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6692..6708 /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(6716..6746) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6761..6782 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6899..6916) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7070..7658) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7150..7169) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7829..568) /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" ORIGIN 1 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 61 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 121 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 181 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 241 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 301 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 361 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 421 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 481 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 541 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 601 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 661 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc 721 gatctcccga tcccctaggg tcgactctca gtacaatctg ctctgatgcc gcatagttaa 781 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt 841 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc 901 gttttgcgct gcttcgctag tcgtgaggct ccggtgcccg tcagtgggca gagcgcacat 961 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa 1021 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg 1081 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt 1141 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg 1201 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc 1261 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt 1321 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt 1381 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat 1441 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc 1501 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca 1561 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc 1621 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg 1681 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc 1741 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac 1801 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt 1861 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag 1921 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag 1981 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat 2041 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt 2101 cgtgaagcgg ccgccaccat ggcccccaag aagaagagga aggtgggcat tcaccgcggg 2161 gtacctatgg tggacttgag gacactcggt tattcgcaac agcaacagga gaaaatcaag 2221 cctaaggtca ggagcaccgt cgcgcaacac cacgaggcgc ttgtggggca tggcttcact 2281 catgcgcata ttgtcgcgct ttcacagcac cctgcggcgc ttgggacggt ggctgtcaaa 2341 taccaagata tgattgcggc cctgcccgaa gccacgcacg aggcaattgt aggggtcggt 2401 aaacagtggt cgggagcgcg agcacttgag gcgctgctga ctgtggcggg tgagcttagg 2461 gggcctccgc tccagctcga caccgggcag ctgctgaaga tcgcgaagag agggggagta 2521 acagcggtag aggcagtgca cgcctggcgc aatgcgctca ccggggcccc cttgaacaga 2581 gacgatctag agctagacgt ctcgctgaca cccgaacagg tggtcgccat tgctagcaac 2641 gggggaggac ggccagcctt ggagagcatt gtggcccagc tgtccaggcc ggaccctgcc 2701 ctggccgcgt taaccaacga ccacctcgtc gccttggcct gcctcggcgg acgtcctgcg 2761 ctggatgcag tgaaaaaggg attgccgcac gcgccggcct tgatcaaaag aaccaatcgc 2821 cgtattcccg aacgcacatc ccatcgcgtt gccgaccacg cgcaagtggt tcgcgtgctg 2881 ggttttttcc agtgccactc ccacccagcg caagcatttg atgacgccat gacgcagttc 2941 gggatgagcg gaggaggatc cctgcccacc tgcagctgtc ttgatcgagt tatacaaaaa 3001 gacaaaggcc catattatac acaccttggg gcaggaccaa gtgttgctgc tgtcagggaa 3061 atcatggaga ataggtatgg tcaaaaagga aacgcaataa ggatagaaat agtagtgtac 3121 accggtaaag aagggaaaag ctctcatggg tgtccaattg ctaagtgggt tttaagaaga 3181 agcagtgatg aagaaaaagt tctttgtttg gtccggcagc gtacaggcca ccactgtcca 3241 actgctgtga tggtggtgct catcatggtg tgggatggca tccctcttcc aatggccgac 3301 cggctataca cagagctcac agagaatcta aagtcataca atgggcaccc taccgacaga 3361 agatgcaccc tcaatgaaaa tcgtacctgt acatgtcaag gaattgatcc agagacttgt 3421 ggagcttcat tctcttttgg ctgttcatgg agtatgtact ttaatggctg taagtttggt 3481 agaagcccaa gccccagaag atttagaatt gatccaagct ctcccttaca tgaaaaaaac 3541 cttgaagata acttacagag tttggctaca cgattagctc caatttataa gcagtatgct 3601 ccagtagctt accaaaatca ggtggaatat gaaaatgttg cccgagaatg tcggcttggc 3661 agcaaggaag gtcgaccctt ctctggggtc actgcttgcc tggacttctg tgctcatccc 3721 cacagggaca ttcacaacat gaataatgga agcactgtgg tttgtacctt aactcgagaa 3781 gataaccgct ctttgggtgt tattcctcaa gatgagcagc tccatgtgct acctctttat 3841 aagctttcag acacagatga gtttggctcc aaggaaggaa tggaagccaa gatcaaatct 3901 ggggccatcg aggtcctggc accccgccgc aaaaaaagaa cgtgtttcac tcagcctgtt 3961 ccccgttctg gaaagaagag ggctgcgatg atgacagagg ttcttgcaca taagataagg 4021 gcagtggaaa agaaacctat tccccgaatc aagcggaaga ataactcaac aacaacaaac 4081 aacagtaagc cttcgtcact gccaacctta gggagtaaca ctgagaccgt gcaacctgaa 4141 gtaaaaagtg aaaccgaacc ccattttatc ttaaaaagtt cagacaacac taaaacttat 4201 tcgctgatgc catccgctcc tcacccagtg aaagaggcat ctccaggctt ctcctggtcc 4261 ccgaagactg cttcagccac accagctcca ctgaagaatg acgcaacagc ctcatgcggg 4321 ttttcagaaa gaagcagcac tccccactgt acgatgcctt cgggaagact cagtggtgcc 4381 aatgctgcag ctgctgatgg ccctggcatt tcacagcttg gcgaagtggc tcctctcccc 4441 accctgtctg ctcctgtgat ggagcccctc attaattctg agccttccac tggtgtgact 4501 gagccgctaa cgcctcatca gccaaaccac cagccctcct tcctcacctc tcctcaagac 4561 cttgcctctt ctccaatgga agaagatgag cagcattctg aagcagatga gcctccatca 4621 gacgaacccc tatctgatga ccccctgtca cctgctgagg agaaattgcc ccacattgat 4681 gagtattggt cagacagtga gcacatcttt ttggatgcaa atattggtgg ggtggccatc 4741 gcacctgctc acggctcggt tttgattgag tgtgcccggc gagagctgca cgctaccact 4801 cctgttgagc accccaaccg taatcatcca acccgcctct cccttgtctt ttaccagcac 4861 aaaaacctaa ataagcccca acatggtttt gaactaaaca agattaagtt tgaggctaaa 4921 gaagctaaga ataagaaaat gaaggcctca gagcaaaaag accaggcagc taatgaaggt 4981 ccagaacagt cctctgaagt aaatgaattg aaccaaattc cttctcataa agcattaaca 5041 ttaacccatg acaatgttgt caccgtgtcc ccttatgctc tcacacacgt tgcggggccc 5101 tataaccatt gggtctgatt aattaattgt ttattgcagc ttataatggt tacaaataaa 5161 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 5221 tgtccaaact catcaatgta tcttatgaat tccgattaag tcgaccctgt ggaatgtgtg 5281 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 5341 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 5401 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 5461 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 5521 ttatgcagag gccgaggccg cctcggcttc tgagctattc cagaagtagt gaggaggctt 5581 ttttggaggc ctaggctttt gcaaaaagct taccatgacc gagtacaagc ccacggtgcg 5641 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga 5701 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct 5761 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga 5821 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc 5881 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat 5941 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg 6001 cgtgtcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga 6061 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc 6121 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gagtgcccga aggaccgcgc 6181 gacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc 6241 gaccgaaagg agcgcacgac cccatgcatc tcgaaggtaa gcctatccct aaccctctcc 6301 tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct 6361 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc 6421 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg 6481 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca 6541 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tccatggctt 6601 ctgaggcgga aagaaccagc tggggctcga taccgtcgac ctctagctag agcttggcgt 6661 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 6721 tacgagccgg aagcataaag tgtaaagcct agggtgccta atgagtgagc taactcacat 6781 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 6841 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 6901 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 6961 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 7021 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 7081 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 7141 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 7201 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 7261 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 7321 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 7381 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 7441 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 7501 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 7561 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 7621 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 7681 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 7741 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 7801 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 7861 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 7921 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 7981 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 8041 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 8101 gtagttcgcc agttaatagt t //