LOCUS       Exported                6519 bp ds-DNA     circular SYN 25-MAY-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    NES-CAGEC-HOXA9c (Npu)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6519)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jun 17, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     LOCUS       Exported               11475 bp ds-DNA     circular SYN 
            10-JAN-2018

            DEFINITION  encodes S. pyogenes dCas9 with c-terminal fusion of 
            human p300 HAT 

                        core (aa 1048-1664) driven by CMV promoter.

            ACCESSION   .

            VERSION     .

            KEYWORDS    pcDNA-dCas9-p300 Core

            SOURCE      synthetic DNA construct

              ORGANISM  synthetic DNA construct

            REFERENCE   1  (bases 1 to 11475)

              AUTHORS   Hilton IB, D'Ippolito AM, Vockley CM, Thakore PI, 
            Crawford GE, Reddy

                        TE, Gersbach CA

              TITLE     Epigenome editing by a CRISPR-Cas9-based 
            acetyltransferase activates

                        genes from promoters and enhancers.

              JOURNAL   Nat Biotechnol. 2015 May;33(5):510-7. doi: 
            10.1038/nbt.3199. Epub 

                        2015 Apr 6.

              PUBMED    25849900

            REFERENCE   2  (bases 1 to 11475)

              AUTHORS   .

              TITLE     Direct Submission

              JOURNAL   Exported Jan 10, 2018 from SnapGene Server 1.1.58

                        http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6519
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable  
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    2092..2113
                     /label=VecF
                     /note="VecF"
     primer_bind     complement(2113..2130)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer.  
                     Also called BGH reverse"
     polyA_signal    2119..2343
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2344..2364)
                     /label=CE-rev sequence
     rep_origin      2389..2817
                     /direction=RIGHT
                     /label=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow  
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2476..2495)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2686..2707
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2826..2846)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2831..3160
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3011..3146
                     /direction=RIGHT
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3073..3092
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3227..4021
                     /codon_start=1
                     /label=aminoglycoside phosphotransferase from
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3281..3300)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3891..3910
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4195..4316
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4232..4251)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4286..4305
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4365..4381)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4365..4381)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar  
                     variants"
     primer_bind     complement(4378..4400)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4389..4405
                     /label=lac operator
                     /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(4413..4443)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4458..4479
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4596..4613)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4767..5352)
                     /direction=LEFT
                     /label=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of  
                     replication"
     primer_bind     complement(4847..4866)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5523..6383)
                     /codon_start=1
                     /label=AmpR (beta-lactamase)
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6146..6165
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6384..6488)
                     /label=bla
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccatggctc tccaaaaaaa gttggaagag cttgaactgg acgaacaggg aggaagtgga
      961 ggtagtctcg agggtaccat gggttcagac tataaagatg atgatgataa aggtggctcc
     1021 gactacaagg atgacgatga caagggaggt agtgactaca aagacgatga cgacaagggc
     1081 ggaggaggaa gtggaggtag tggaagagta gcgatcctct ggcatgagat gtggcatgaa
     1141 ggcctggaag aggcatctcg tttgtacttt ggggaaagga acgtgaaagg catgtttgag
     1201 gtgctggagc ccttgcatgc tatgatggaa cggggccccc agactctgaa ggaaacatcc
     1261 tttaatcagg cctatggtcg agatttaatg gaggcccaag agtggtgcag gaagtacatg
     1321 aaatcaggga atgtcaagga cctcctccaa gcctgggacc tctattatca tgtgttccga
     1381 cgaatctcag gaaatggaaa tggaggagaa caagaggtgt ttgagtattg tttggaagat
     1441 ggttcattga ttcgggcaac aaaagaccat aagtttatga ctgttgatgg tcaaatgttg
     1501 ccaattgatg aaatatttga acgtgaattg gatttgatgc gggttgataa tttgccgaat
     1561 ggaagtggag gagagaacct gtattttcag ggcgagaacc tgtattttca gggcggaagt
     1621 ggaggaatca aaatagccac acgtaaatat ttaggcaaac aaaatgtcta tgacattgga
     1681 gttgagcgcg accataattt tgcactcaaa aatggcttca tagcttctaa ttgctttaat
     1741 agagaaaaac aacccagcga aggcgccttc tctgaaaaca atgctgagaa tgagagcggc
     1801 ggagacaagc cccccatcga tcccaataac ccagcagcca actggcttca tgcgcgctcc
     1861 actcggaaaa agcggtgccc ctatacaaaa caccagaccc tggaactgga gaaagagttt
     1921 ctgttcaaca tgtacctcac cagggaccgc aggtacgagg tggctcgact gctcaacctc
     1981 accgagaggc aggtcaagat ctggttccag aaccgcagga tgaaaatgaa gaaaatcaac
     2041 aaagaccgag caaaagacga gtacccgtac gacgttccgg actacgcttc ttgagtttaa
     2101 accgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2161 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2221 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     2281 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     2341 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     2401 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2461 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2521 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2581 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2641 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2701 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     2761 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     2821 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2881 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2941 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3001 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3061 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     3121 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     3181 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3241 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3301 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3361 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3421 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3481 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3541 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3601 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3661 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3721 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     3781 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3841 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3901 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3961 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4021 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4081 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4141 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     4201 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4261 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4321 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     4381 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4441 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4501 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4561 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4621 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4681 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4741 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4801 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4861 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4921 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4981 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5041 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5101 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5161 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5221 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5281 aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5341 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5401 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5461 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5521 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5581 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5641 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5701 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5761 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5821 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5881 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5941 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6001 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6061 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6121 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6181 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6241 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6301 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6361 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6421 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6481 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
