LOCUS       sequence-194598-        7452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dSa Cas9 fused to optimized combination of truncated 
            activation domains from SCP1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7452)
  AUTHORS   Vora S, Cheng J, Xiao R, VanDusen NJ, Quintino L, Pu WT, 
            Vandenberghe LH, Chavez A, Church G
  TITLE     Rational design of a compact CRISPR-Cas9 activator for AAV-mediated 
            delivery
  JOURNAL   bioRxiv 2018. 298620
REFERENCE   2  (bases 1 to 7452)
  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: "bioRxiv 
            2018. 298620"
COMMENT     Sequence Label: pAAV-SCP1-dSa-VPR mini.
FEATURES             Location/Qualifiers
     source          1..7452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5..27)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     127..146
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(163..618)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(300..321)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     512..531
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   693..833
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   704..833
                     /label=AAV2 ITR
     regulatory      935..944
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             947..967
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4148..4195
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4226..4375
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             4400..4420
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4580..4936
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     primer_bind     complement(5192..5209)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5198..5405
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5418..5547
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(5609..6197)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5689..5708)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6368..7159,7160..7228))
                     /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"
     primer_bind     6991..7010
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7229..7333)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7401..7419
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
       61 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
      121 actatgcggc atcagagcag attgtactga gagtgcacca taaaattgta aacgttaata
      181 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caatagaccg
      241 aaatcggcaa aatcccttat aaatcaaaag aatagcccga gatagagttg agtgttgttc
      301 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
      361 ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt
      421 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
      481 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
      541 aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
      601 cgccgctaca gggcgcgtac tatggttgct ttgacgtatg cggtgtgaaa taccgcacag
      661 atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct
      721 cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
      781 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
      841 cctcgaggta cttatataag ggggtggggg cgcgttcgtc ctcagtcgcg atcgaacact
      901 cgagccgagc agacgtgcct acggaccgac cggtgccacc atggccccaa agaagaagcg
      961 gaaggtcggt atccacggag tcccagcagc caagcggaac tacatcctgg gcctggccat
     1021 cggcatcacc agcgtgggct acggcatcat cgactacgag acacgggacg tgatcgatgc
     1081 cggcgtgcgg ctgttcaaag aggccaacgt ggaaaacaac gagggcaggc ggagcaagag
     1141 aggcgccaga aggctgaagc ggcggaggcg gcatagaatc cagagagtga agaagctgct
     1201 gttcgactac aacctgctga ccgaccacag cgagctgagc ggcatcaacc cctacgaggc
     1261 cagagtgaag ggcctgagcc agaagctgag cgaggaagag ttctctgccg ccctgctgca
     1321 cctggccaag agaagaggcg tgcacaacgt gaacgaggtg gaagaggaca ccggcaacga
     1381 gctgtccacc aaagagcaga tcagccggaa cagcaaggcc ctggaagaga aatacgtggc
     1441 cgaactgcag ctggaacggc tgaagaaaga cggcgaagtg cggggcagca tcaacagatt
     1501 caagaccagc gactacgtga aagaagccaa acagctgctg aaggtgcaga aggcctacca
     1561 ccagctggac cagagcttca tcgacaccta catcgacctg ctggaaaccc ggcggaccta
     1621 ctatgaggga cctggcgagg gcagcccctt cggctggaag gacatcaaag aatggtacga
     1681 gatgctgatg ggccactgca cctacttccc cgaggaactg cggagcgtga agtacgccta
     1741 caacgccgac ctgtacaacg ccctgaacga cctgaacaat ctcgtgatca ccagggacga
     1801 gaacgagaag ctggaatatt acgagaagtt ccagatcatc gagaacgtgt tcaagcagaa
     1861 gaagaagccc accctgaagc agatcgccaa agaaatcctc gtgaacgaag aggatattaa
     1921 gggctacaga gtgaccagca ccggcaagcc cgagttcacc aacctgaagg tgtaccacga
     1981 catcaaggac attaccgccc ggaaagagat tattgagaac gccgagctgc tggatcagat
     2041 tgccaagatc ctgaccatct accagagcag cgaggacatc caggaagaac tgaccaatct
     2101 gaactccgag ctgacccagg aagagatcga gcagatctct aatctgaagg gctataccgg
     2161 cacccacaac ctgagcctga aggccatcaa cctgatcctg gacgagctgt ggcacaccaa
     2221 cgacaaccag atcgctatct tcaaccggct gaagctggtg cccaagaagg tggacctgtc
     2281 ccagcagaaa gagatcccca ccaccctggt ggacgacttc atcctgagcc ccgtcgtgaa
     2341 gagaagcttc atccagagca tcaaagtgat caacgccatc atcaagaagt acggcctgcc
     2401 caacgacatc attatcgagc tggcccgcga gaagaactcc aaggacgccc agaaaatgat
     2461 caacgagatg cagaagcgga accggcagac caacgagcgg atcgaggaaa tcatccggac
     2521 caccggcaaa gagaacgcca agtacctgat cgagaagatc aagctgcacg acatgcagga
     2581 aggcaagtgc ctgtacagcc tggaagccat ccctctggaa gatctgctga acaacccctt
     2641 caactatgag gtggaccaca tcatccccag aagcgtgtcc ttcgacaaca gcttcaacaa
     2701 caaggtgctc gtgaagcagg aagaagccag caagaagggc aaccggaccc cattccagta
     2761 cctgagcagc agcgacagca agatcagcta cgaaaccttc aagaagcaca tcctgaatct
     2821 ggccaagggc aagggcagaa tcagcaagac caagaaagag tatctgctgg aagaacggga
     2881 catcaacagg ttctccgtgc agaaagactt catcaaccgg aacctggtgg ataccagata
     2941 cgccaccaga ggcctgatga acctgctgcg gagctacttc agagtgaaca acctggacgt
     3001 gaaagtgaag tccatcaatg gcggcttcac cagctttctg cggcggaagt ggaagtttaa
     3061 gaaagagcgg aacaaggggt acaagcacca cgccgaggac gccctgatca ttgccaacgc
     3121 cgatttcatc ttcaaagagt ggaagaaact ggacaaggcc aaaaaagtga tggaaaacca
     3181 gatgttcgag gaaaagcagg ccgagagcat gcccgagatc gaaaccgagc aggagtacaa
     3241 agagatcttc atcacccccc accagatcaa gcacattaag gacttcaagg actacaagta
     3301 cagccaccgg gtggacaaga agcctaatag agagctgatt aacgacaccc tgtactccac
     3361 ccggaaggac gacaagggca acaccctgat cgtgaacaat ctgaacggcc tgtacgacaa
     3421 ggacaatgac aagctgaaaa agctgatcaa caagagcccc gaaaagctgc tgatgtacca
     3481 ccacgacccc cagacctacc agaaactgaa gctgattatg gaacagtacg gcgacgagaa
     3541 gaatcccctg tacaagtact acgaggaaac cgggaactac ctgaccaagt actccaaaaa
     3601 ggacaacggc cccgtgatca agaagattaa gtattacggc aacaaactga acgcccatct
     3661 ggacatcacc gacgactacc ccaacagcag aaacaaggtc gtgaagctgt ccctgaagcc
     3721 ctacagattc gacgtgtacc tggacaatgg cgtgtacaag ttcgtgaccg tgaagaatct
     3781 ggatgtgatc aaaaaagaaa actactacga agtgaatagc aagtgctatg aggaagctaa
     3841 gaagctgaag aagatcagca accaggccga gtttatcgcc tccttctaca acaacgatct
     3901 gatcaagatc aacggcgagc tgtatagagt gatcggcgtg aacaacgacc tgctgaaccg
     3961 gatcgaagtg aacatgatcg acatcaccta ccgcgagtac ctggaaaaca tgaacgacaa
     4021 gaggcccccc aggatcatta agacaatcgc ctccaagacc cagagcatta agaagtacag
     4081 cacagacatt ctgggcaacc tgtatgaagt gaaatctaag aagcaccctc agatcatcaa
     4141 aaagggcaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga aaaagggatc
     4201 cgaggccagc ggttccggac gggctgacgc attggacgat tttgatctgg atatgctggg
     4261 aagtgacgcc ctcgatgatt ttgaccttga catgcttggt tcggatgccc ttgatgactt
     4321 tgacctcgac atgctcggca gtgacgccct tgatgatttc gacctggaca tgctgattaa
     4381 ctctagaagt tccggatctc cgaaaaagaa acgcaaagtt ggttcgggag gtggttcggg
     4441 tggctctgga tcagtgctgc ctcaggctcc tgctcctgca ccagctccag ccatggtgtc
     4501 tgcactggct caggcaccag cacccgtgcc tgtgctggct cctggacctc cacaggctgt
     4561 ggctccacca gcccctaaac ctacacaggc cggcgagggc acactgtctg aagctctgct
     4621 gcagctgcag ttcgacgacg aggatctggg agccctgctg ggaaacagca ccgatcctgc
     4681 cgtgttcacc gacctggcca gcgtggacaa cagcgagttc cagcagctgc tgaaccaggg
     4741 catccctgtg gcccctcaca ccaccgagcc catgctgatg gaataccccg aggccatcac
     4801 ccggctcgtg acaggcgctc agaggcctcc tgatccagct cctgcccctc tgggagcacc
     4861 aggcctgcct aatggactgc tgtctggcga cgaggacttc agctctatcg ccgatatgga
     4921 tttctcagcc ttgctgtcag gcggtggtag tggtgggagc ggtagtgacc tttcccatcc
     4981 gcccccaagg ggccatctgg atgagctgac aaccacactt gagtccatga ccgaggatct
     5041 gaacctggac tcacccctga ccccggaatt gaacgagatt ctggatacct tcctgaacga
     5101 cgagtgcctc ttgcatgcca tgcatatcag cacaggactg tccatcttcg acacatctct
     5161 gttttaggaa ttcctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     5221 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     5281 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     5341 gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag caggcatgct
     5401 ggggagctag aggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     5461 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga
     5521 gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga
     5581 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5641 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5701 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5761 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5821 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5881 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5941 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6001 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6061 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6121 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6181 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6241 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6301 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6361 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6421 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6481 ctggccccag tgctgcaatg ataccgcggc ttccacgctc accggctcca gatttatcag
     6541 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6601 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6661 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6721 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6781 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6841 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6901 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6961 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7021 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7081 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7141 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7201 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7261 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7321 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     7381 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     7441 gtgatgacgg tg
//
