LOCUS       Exported                8547 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 8547)
  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 8547)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8547
                     /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        926..2104
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1157..2095
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2052..2072
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      2112..2121
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2124..2144
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5589..5608)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5643..5662
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5699..5719)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5704..6033
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5884..6019
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5946..5965
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6043..6642
                     /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(6043..6062)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6539..6559
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             6702..6743
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6753..6770
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6793..6810)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6799..7023
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7094..7110)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7094..7110)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7107..7129)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7118..7134
                     /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(7142..7172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7187..7208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7325..7342)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7496..8084)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7576..7595)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8255..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 gcttcgtagt cgtgaggctc cggtgcccgt cagtgggcag agcgcacatc
      961 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
     1021 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     1081 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     1141 tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg
     1201 ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc
     1261 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
     1321 gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
     1381 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
     1441 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca
     1501 cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac
     1561 atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca
     1621 agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
     1681 ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc
     1741 tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc
     1801 cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta
     1861 ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg
     1921 ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
     1981 taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc
     2041 ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc
     2101 gtgaagcggc cgccaccatg gcccccaaga agaagaggaa ggtgggcatt caccgcgggg
     2161 tacctatggt ggacttgagg acactcggtt attcgcaaca gcaacaggag aaaatcaagc
     2221 ctaaggtcag gagcaccgtc gcgcaacacc acgaggcgct tgtggggcat ggcttcactc
     2281 atgcgcatat tgtcgcgctt tcacagcacc ctgcggcgct tgggacggtg gctgtcaaat
     2341 accaagatat gattgcggcc ctgcccgaag ccacgcacga ggcaattgta ggggtcggta
     2401 aacagtggtc gggagcgcga gcacttgagg cgctgctgac tgtggcgggt gagcttaggg
     2461 ggcctccgct ccagctcgac accgggcagc tgctgaagat cgcgaagaga gggggagtaa
     2521 cagcggtaga ggcagtgcac gcctggcgca atgcgctcac cggggccccc ttgaacagag
     2581 acgatctaga gctagacgtc tcgctgacac ccgaacaggt ggtcgccatt gcttcccacg
     2641 acggaggacg gccagccttg gagagcattg tggcccagct gtccaggccg gaccctgccc
     2701 tggccgcgtt aaccaacgac cacctcgtcg ccttggcctg cctcggcgga cgtcctgcgc
     2761 tggatgcagt gaaaaaggga ttgccgcacg cgccggcctt gatcaaaaga accaatcgcc
     2821 gtattcccga acgcacatcc catcgcgttg ccgaccacgc gcaagtggtt cgcgtgctgg
     2881 gttttttcca gtgccactcc cacccagcgc aagcatttga tgacgccatg acgcagttcg
     2941 ggatgagcgg aggaggatct ttatctggga agaaggcggc agccgcggcg gcggcggctg
     3001 cagcggcagc aaccgggacg gaggctggcc ctgggacagc aggcggctcc gagaacgggt
     3061 ctgaggtggc cgcgcagccc gcgggcctgt cgggcccagc cgaggtcggg ccgggggcgg
     3121 tgggggagcg cacaccccgc aagaaagagc ctccgcgggc ctcgcccccc gggggcctgg
     3181 cggaaccgcc ggggtccgca gggcctcagg ccggccctac tgtcgtgcct gggtctgcga
     3241 cccccatgga aactggaata gcagagactc cggaggggcg tcggaccagc cggcgcaagc
     3301 gggcgaaggt agagtacaga gagatggatg aaagcttggc caacctctca gaagatgagt
     3361 attattcaga agaagagaga aatgccaaag cagagaagga aaagaagctt cccccaccac
     3421 cccctcaagc cccacctgag gaagaaaatg aaagtgagcc tgaagaacca tcgggtgtgg
     3481 agggcgcagc tttccagagc cgacttcctc atgaccggat gacttctcaa gaagcagcct
     3541 gttttccaga tattatcagt ggaccacaac agacccagaa ggtttttctt ttcattagaa
     3601 accgcacact gcagttgtgg ttggataatc caaagattca gctgacattt gaggctactc
     3661 tccaacaatt agaagcacct tataacagtg atactgtgct tgtccaccga gttcacagtt
     3721 atttagagcg tcatggtctt atcaacttcg gcatctataa gaggataaaa cccctaccaa
     3781 ctaaaaagac aggaaaggta attattatag gctctggggt ctcaggcttg gcagcagctc
     3841 gacagttaca aagttttgga atggatgtca cacttttgga agccagggat cgtgtgggtg
     3901 gacgagttgc cacatttcgc aaaggaaact atgtagctga tcttggagcc atggtggtaa
     3961 caggtcttgg agggaatcct atggctgtgg tcagcaaaca agtaaatatg gaactggcca
     4021 agatcaagca aaaatgccca ctttatgaag ccaacggaca agctgttcct aaagagaaag
     4081 atgaaatggt agagcaagag tttaaccggt tgctagaagc tacatcttac cttagtcatc
     4141 aactagactt caatgtcctc aataataagc atgtgtccct tggccaggca ttggaagttg
     4201 tcattcagtt acaagagaag catgtcaaag atgagcagat tgaacattgg aagaagatag
     4261 tgaaaactca ggaagaattg aaagaacttc ttaataagat ggtaaatttg aaagagaaaa
     4321 ttaaagaact ccatcagcaa tacaaagaag catctgaagt aaagccaccc agagatatta
     4381 ctgccgagtt cttagtgaaa agcaaacaca gggatctgac cgccctatgc aaggaatatg
     4441 atgaattagc tgaaacacaa ggaaagctag aagaaaaact tcaggagttg gaagcgaatc
     4501 ccccaagtga tgtatatctc tcatcaagag acagacaaat acttgattgg cattttgcaa
     4561 atcttgaatt tgctaatgcc acacctctct caactctctc ccttaagcac tgggatcagg
     4621 atgatgactt tgagttcact ggcagccacc tgacagtaag gaatggctac tcgtgtgtgc
     4681 ctgtggcttt agcagaaggc ctagacatta aactgaatac agcagtgcga caggttcgct
     4741 acacggcttc aggatgtgaa gtgatagctg tgaatacccg ctccacgagt caaaccttta
     4801 tttataaatg cgacgcagtt ctctgtaccc ttcccctggg tgtgctgaag cagcagccac
     4861 cagccgttca gtttgtgcca cctctccctg agtggaaaac atctgcagtc caaaggatgg
     4921 gatttggcaa ccttaacaag gtggtgttgt gttttgatcg ggtgttctgg gatccaagtg
     4981 tcaatttgtt cgggcatgtt ggcagtacga ctgccagcag gggtgagctc ttcctcttct
     5041 ggaacctcta taaagctcca atactgttgg cactagtggc aggagaagct gctggtatca
     5101 tggaaaacat aagtgacgat gtgattgttg gccgatgcct ggccattctc aaagggattt
     5161 ttggtagcag tgcagtacct cagcccaaag aaactgtggt gtctcgttgg cgtgctgatc
     5221 cctgggctcg gggctcttat tcctatgttg ctgcaggatc atctggaaat gactatgatt
     5281 taatggctca gccaatcact cctggcccct cgattccagg tgccccacag ccgattccac
     5341 gactcttctt tgcgggagaa catacgatcc gtaactaccc agccacagtg catggtgctc
     5401 tgctgagtgg gctgcgagaa gcgggaagaa ttgcagacca gtttttgggg gccatgtata
     5461 cgctgcctcg ccaggccaca ccaggtgttc ctgcacagca gtccccaagc atgtgagaca
     5521 gatgcattct aagggaagag gcccatttaa ttaattgttt attgcagctt ataatggtta
     5581 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5641 ttgtggtttg tccaaactca tcaatgtatc ttatgaattc cgattaagtc gaccctgtgg
     5701 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5761 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5821 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5881 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5941 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     6001 ggaggctttt ttggaggcct aggcttttgc aaaaagctta ccatgaccga gtacaagccc
     6061 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     6121 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     6181 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     6241 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     6301 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     6361 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     6421 accgtcggcg tgtcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     6481 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     6541 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga gtgcccgaag
     6601 gaccgcgcga cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg
     6661 cagcgcccga ccgaaaggag cgcacgaccc catgcatcga aggtaagcct atccctaacc
     6721 ctctcctcgg tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa
     6781 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     6841 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     6901 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     6961 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     7021 tggcttctga ggcggaaaga accagctggg gctcgatacc gtcgacctct agctagagct
     7081 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     7141 acaacatacg agccggaagc ataaagtgta aagcctaggg tgcctaatga gtgagctaac
     7201 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     7261 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     7321 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7381 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     7441 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     7501 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     7561 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7621 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7681 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7741 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7801 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7861 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7921 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7981 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     8041 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     8101 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     8161 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     8221 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     8281 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8341 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8401 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8461 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8521 gagtaagtag ttcgccagtt aatagtt
//
