LOCUS Exported 8550 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Backbone contains SHD/C 0.5 Domain TALE N-terminal: TAL C-terminus: LSD1: Isoform 2. ACCESSION . VERSION . KEYWORDS EMM65 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8550) AUTHORS Mendenhall EM, Williamson KE, Reyon D, Zou JY, Ram O, Joung JK, Bernstein BE TITLE Locus-specific editing of histone modifications at endogenous enhancers. JOURNAL Nat Biotechnol. 2013 Sep 8. doi: 10.1038/nbt.2701. PUBMED 24013198 REFERENCE 2 (bases 1 to 8550) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8550 /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(5590..5609) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5644..5663 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5700..5720) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5705..6034 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5885..6020 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5947..5966 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6044..6643 /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(6044..6063) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 6540..6560 /label=Puro-F /note="Puromycin resistance gene, forward primer" CDS 6705..6746 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 6756..6773 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(6796..6813) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6802..7026 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(7097..7113) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7097..7113) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7110..7132) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7121..7137 /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(7145..7175) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7190..7211 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7328..7345) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7499..8087) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7579..7598) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8258..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 tgcttcccac 2641 gacggaggac 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 tttatctggg aagaaggcgg cagccgcggc ggcggcggct 3001 gcagcggcag caaccgggac ggaggctggc cctgggacag caggcggctc cgagaacggg 3061 tctgaggtgg ccgcgcagcc cgcgggcctg tcgggcccag ccgaggtcgg gccgggggcg 3121 gtgggggagc gcacaccccg caagaaagag cctccgcggg cctcgccccc cgggggcctg 3181 gcggaaccgc cggggtccgc agggcctcag gccggcccta ctgtcgtgcc tgggtctgcg 3241 acccccatgg aaactggaat agcagagact ccggaggggc gtcggaccag ccggcgcaag 3301 cgggcgaagg tagagtacag agagatggat gaaagcttgg ccaacctctc agaagatgag 3361 tattattcag aagaagagag aaatgccaaa gcagagaagg aaaagaagct tcccccacca 3421 ccccctcaag ccccacctga ggaagaaaat gaaagtgagc ctgaagaacc atcgggtgtg 3481 gagggcgcag ctttccagag ccgacttcct catgaccgga tgacttctca agaagcagcc 3541 tgttttccag atattatcag tggaccacaa cagacccaga aggtttttct tttcattaga 3601 aaccgcacac tgcagttgtg gttggataat ccaaagattc agctgacatt tgaggctact 3661 ctccaacaat tagaagcacc ttataacagt gatactgtgc ttgtccaccg agttcacagt 3721 tatttagagc gtcatggtct tatcaacttc ggcatctata agaggataaa acccctacca 3781 actaaaaaga caggaaaggt aattattata ggctctgggg tctcaggctt ggcagcagct 3841 cgacagttac aaagttttgg aatggatgtc acacttttgg aagccaggga tcgtgtgggt 3901 ggacgagttg ccacatttcg caaaggaaac tatgtagctg atcttggagc catggtggta 3961 acaggtcttg gagggaatcc tatggctgtg gtcagcaaac aagtaaatat ggaactggcc 4021 aagatcaagc aaaaatgccc actttatgaa gccaacggac aagctgttcc taaagagaaa 4081 gatgaaatgg tagagcaaga gtttaaccgg ttgctagaag ctacatctta ccttagtcat 4141 caactagact tcaatgtcct caataataag catgtgtccc ttggccaggc attggaagtt 4201 gtcattcagt tacaagagaa gcatgtcaaa gatgagcaga ttgaacattg gaagaagata 4261 gtgaaaactc aggaagaatt gaaagaactt cttaataaga tggtaaattt gaaagagaaa 4321 attaaagaac tccatcagca atacaaagaa gcatctgaag taaagccacc cagagatatt 4381 actgccgagt tcttagtgaa aagcaaacac agggatctga ccgccctatg caaggaatat 4441 gatgaattag ctgaaacaca aggaaagcta gaagaaaaac ttcaggagtt ggaagcgaat 4501 cccccaagtg atgtatatct ctcatcaaga gacagacaaa tacttgattg gcattttgca 4561 aatcttgaat ttgctaatgc cacacctctc tcaactctct cccttaagca ctgggatcag 4621 gatgatgact ttgagttcac tggcagccac ctgacagtaa ggaatggcta ctcgtgtgtg 4681 cctgtggctt tagcagaagg cctagacatt aaactgaata cagcagtgcg acaggttcgc 4741 tacacggctt caggatgtga agtgatagct gtgaataccc gctccacgag tcaaaccttt 4801 atttataaat gcgacgcagt tctctgtacc cttcccctgg gtgtgctgaa gcagcagcca 4861 ccagccgttc agtttgtgcc acctctccct gagtggaaaa catctgcagt ccaaaggatg 4921 ggatttggca accttaacaa ggtggtgttg tgttttgatc gggtgttctg ggatccaagt 4981 gtcaatttgt tcgggcatgt tggcagtacg actgccagca ggggtgagct cttcctcttc 5041 tggaacctct ataaagctcc aatactgttg gcactagtgg caggagaagc tgctggtatc 5101 atggaaaaca taagtgacga tgtgattgtt ggccgatgcc tggccattct caaagggatt 5161 tttggtagca gtgcagtacc tcagcccaaa gaaactgtgg tgtctcgttg gcgtgctgat 5221 ccctgggctc ggggctctta ttcctatgtt gctgcaggat catctggaaa tgactatgat 5281 ttaatggctc agccaatcac tcctggcccc tcgattccag gtgccccaca gccgattcca 5341 cgactcttct ttgcgggaga acatacgatc cgtaactacc cagccacagt gcatggtgct 5401 ctgctgagtg ggctgcgaga agcgggaaga attgcagacc agtttttggg ggccatgtat 5461 acgctgcctc gccaggccac accaggtgtt cctgcacagc agtccccaag catgtgagac 5521 agatgcattc taagggaaga ggcccattta attaattgtt tattgcagct tataatggtt 5581 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 5641 gttgtggttt gtccaaactc atcaatgtat cttatgaatt ccgattaagt cgaccctgtg 5701 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 5761 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg 5821 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc 5881 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccctatg gctgactaat 5941 tttttttatt tatgcagagg ccgaggccgc ctcggcttct gagctattcc agaagtagtg 6001 aggaggcttt tttggaggcc taggcttttg caaaaagctt accatgaccg agtacaagcc 6061 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc 6121 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt 6181 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt 6241 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc 6301 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca 6361 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc 6421 caccgtcggc gtgtcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc 6481 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg 6541 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg agtgcccgaa 6601 ggaccgcgcg acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc 6661 gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg cgaaggtaag cctatcccta 6721 accctctcct cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt 6781 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct 6841 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg 6901 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc 6961 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct 7021 ctatggcttc tgaggcggaa agaaccagct ggggctcgat accgtcgacc tctagctaga 7081 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 7141 cacacaacat acgagccgga agcataaagt gtaaagccta gggtgcctaa tgagtgagct 7201 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 7261 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 7321 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 7381 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 7441 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 7501 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 7561 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 7621 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 7681 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 7741 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 7801 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 7861 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 7921 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct 7981 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 8041 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 8101 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 8161 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 8221 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 8281 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 8341 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 8401 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 8461 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 8521 ctagagtaag tagttcgcca gttaatagtt //