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    pJFA345D4
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 tgctaataat
     2641 aacggaggac 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
//
