LOCUS       sequence-72027-e        8550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone contains SHD/C 0.5 Domain TALE N-terminal: TAL C-terminus: 
            LSD1: Isoform 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2013 Sep 8. doi: 10.1038/nbt.2701."
COMMENT     Sequence Label: EMM65
FEATURES             Location/Qualifiers
     source          1..8550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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"
     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(join(7145..7151,7152..7169,7170..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(join(8258..8550,1..499,500..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
//
