LOCUS       Exported                4416 bp ds-DNA     circular SYN 29-AUG-2018
DEFINITION  CMV-T7 promoter expression plasmid for human codon optimized AcrVA2 
            anti-CRISPR protein with C-terminal NLS  .
ACCESSION   urn.local...e-9a99gh3
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4416)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 29, 2018 from SnapGene 4.2.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4416
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1..3
                     /label=start codon
                     /note="/Color=#ff7777"
                     /note="/created_by=User"
                     /note="/modified_by=User"
     CDS             4..969
                     /label=human codon optimized AcrVA2
                     /note="/created_by=User"
                     /note="/modified_by=User"
     CDS             4..6
                     /label=glycine added for Kozak
                     /note="/Color=#ff7777"
                     /note="/created_by=User"
                     /note="/modified_by=User"
     CDS             970..993
                     /label=GS linker
                     /note="/Color=#ff7400"
                     /note="/created_by=User"
                     /note="/modified_by=User"
     CDS             994..1014
                     /label=NLS(SV40)
                     /note="/Color=#6bba70"
                     /note="/created_by=User"
     terminator      1066..1293
                     /label=bGH PA terminator
                     /note="/modified_by=User"
                     /note="/translation=MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTD
                     RHSIK 
                     KNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLE 
                     ESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAH 
                     MIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSK 
                     SRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLD 
                     NLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTL 
                     LKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVK 
                     LNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPY 
                     YVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVL 
                     PKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKE 
                     DYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLT 
                     LFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDF 
                     LKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQT 
                     VKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKE 
                     HPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKV 
                     LTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKA 
                     GFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF 
                     YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIG 
                     KATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS 
                     MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVV 
                     AKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLF 
                     ELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQH 
                     KHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGA 
                     PAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDGSPKKKRKVS 
                     SDYKDHDGDYKDHDIDYKDDDDKAAG*"
     misc_feature    complement(1382..1409)
                     /label=lacO reg
                     /note="Geneious type: regulatory sequence"
                     /note="/Imported_from=Pla sMapper"
                     /note="/Transferred_From=pCAG-hCas9-nls-3XFLAG (SQT817), 
                     pCAG-hCas9-nls-3XFLAG delta NcoI (BPK742), pCMV-T7-Cas9 
                     (MLM3613), pCMV-T7-Cas9 delta NcoI"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=lacO reg; 
                     pCAG-Cas9-RECODED-nls-3XFLAG.xdna (SQT817); 
                     pCAG-hCas9(R1333A)-NLS-3xFLAG; 
                     pCAG-hCas9(R1333A/R1335A)-NLS-3xFLAG; 
                     pCAG-hCas9(R1333Q)-NLS-3xFLAG; 
                     pCAG-hCas9(R1333Q/R1335Q)-NLS-3xFLAG; 
                     pCAG-hCas9(R1335A)-NLS-3xFLAG; 
                     pCAG-hCas9(R1335Q)-NLS-3xFLAG; pCAG-hCas9-NLS-3xFLAG 
                     (SQT817)"
                     /note="/X-Transferred_Similarity=100.00%; 89.29%"
     promoter        complement(1419..1442)
                     /label=lac promoter
                     /note="/Transferred_From=pCMV-T7-Cas9 (MLM3613), 
                     pCMV-T7-Cas9 delta NcoI"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=JDS246; 
                     VC-(VC780+Csy4-Cas9)_pLKO-TRC001; pAC90-pmax-DEST (Addgene 
                     48222); pCAG-DEST (BPK88); pCAG-DEST V2; 
                     pCMV-NLS-hTtAgo-NLS-3xFLAG; pCW-Cas9 (modified); pCW-Cas9 
                     Activator; pCW-Cas9 N-F-N Activator; pCW-Cas9 N-F-N 
                     Activator-GG; pCW-EFS-Puro-P2A-NFN-dCas9-VP64; 
                     pCW-NLS-dCas9-VP64 EFS-Puro-T2A-rTta3-WPRE; pINDUCER21 
                     (ORF-EG) (modified); pInducer20 (modified); 
                     pLV-CMVEI.EGFPDS-iNpLTRSEBBA6_0693; pLenti CMV Puro DEST 
                     (w118-1) (Addgene 17452); pLenti X1 Puro DEST (694-6) 
                     (Addgene 17297); pLenti-CAG-Puro-DEST; 
                     pLenti-TREminCMV-NLS-3xFLAG-NLS-dCas9-VP64 
                     EFS-Puro-T2A-rTta3-WPRE; pLenti-TREminCMV-NLS-dCas9-VP64 
                     EFS-Puro-T2A-rTta3-WPRE; 
                     pLenti-U6-DEST-EF1a-Csy4-P2A-Neo-WPRE; 
                     pLenti-U6-DEST-EF1a-Csy4-P2A-hygro; 
                     pLenti-U6-DEST-EF1a-Csy4-P2A-hygro-WPRE; 
                     pLenti-U6-DEST-EFS-Csy4-P2A-hygro; 
                     pLenti-U6-DEST-EFS-Csy4-P2A-hygro-WPRE; 
                     pLenti-U6-DEST-SpeI/NheI-P2A-Puro-WPRE; 
                     pLenti-U6-DEST-strategy2cassette; pLenti-U6-Puro-DEST 
                     (BPK162); pLenti-U6-Puro-DEST + WPRE (BPK640); 
                     pLenti-U6-Puro-DEST-WPRE; pMLM3705; pMLM3705 annotation 
                     mess; pUC19-U6-DEST (BPK84); pcDNA5/FRT; 
                     pcDNA5/FRT-tdtomato-AAVS1 site 1; pcDNA5/FRT-tdtomato-AAVS1
                     site 1 in wrong way; pcDNA5/FRT-tdtomato-AAVS1 site 2"
                     /note="/X-Transferred_Similarity=100.00%"
                     /note="/X-Transferred_Translation=MAVFIHNFKRKGGIGGYSAGERIVD
                     IIATD 
                     IQTKELQKQITKIQNFRVYYRDSRDPVWTRILYHSLSVIEKSESRVYHSLSVIEKSES 
                     RVYHSLSVIEKSESRVYHSLSVIEKSESRVYHSLSVIEKSESRVYHSLSVIEKSESRV 
                     YHSLSVIEKSESRARYPGRGRRVRWEAYISRARLVNRQIAWRRHPRCFDLHRRHRDRS 
                     SLRGPELVQCGGILQISTSLYKKAERET*; 
                     MEWDREINNYTSLIHSLIEESQNQQEKNEQELLELDKWASLWNWFNITNWLWYIKLFI 
                     MIVGGLVGLRIVFAVLSIVNRVRQGYSPLSFQTHLPTPRGPDRPEGIEEEGGERDRDR 
                     SIRLVNGSRRLTFKRKGGIGGYSAGERIVDIIATDIQTKELQKQITKIQNFIDKLGSS 
                     ALHNLR*; 
                     MEWDREINNYTSLIHSLIEESQNQQEKNEQELLELDKWASLWNWFNITNWLWYIKLFI 
                     MIVGGLVGLRIVFAVLSIVNRVRQGYSPLSFQTHLPTPRGPDRPEGIEEEGGERDRDR 
                     SIRLVNGSRRLTFKRKGGIGGYSAGERIVDIIATDIQTKELQKQITKIQNFIPVWWNS 
                     ADINKFVQKS*"
                     /note="/modified_by=User"
     protein_bind    complement(1462..1474)
                     /label=catabolite activator protein (CAP) binding site
                     /bound_moiety="catabolite activator protein (CAP)"
                     /note="CAP protein binding site"
                     /note="/Transferred_From=pCMV-T7-Cas9 (MLM3613), 
                     pCMV-T7-Cas9 delta NcoI"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=pNEB193; pNEB205A; pNEB206A; 
                     pUC19"
                     /note="/X-Transferred_Similarity=100.00%"
     terminator      1552..1683
                     /label=SV40 PA terminator
                     /note="/Transferred_From=pCMV-T7-Cas9 (MLM3613), 
                     pCMV-T7-Cas9 delta NcoI"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_Similarity=100.00%"
                     /note="/modified_by=User"
     misc_feature    complement(1751..2370)
                     /label=pBR322 origin
                     /note="Geneious type: origin of replication"
                     /note="/Imported_from=Pla sMapper"
                     /note="/Transferred_From=pCAG-hCas9-nls-3XFLAG (SQT817), 
                     pCAG-hCas9-nls-3XFLAG delta NcoI (BPK742), pCMV-T7-Cas9 
                     (MLM3613), pCMV-T7-Cas9 delta NcoI"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=pBR322 origin"
                     /note="/X-Transferred_Similarity=100.00%"
     gene            complement(2525..3385)
                     /gene="Ampicillin"
                     /label=Ampicillin
                     /note="/translation=MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTD
                     RHSIK 
                     KNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLE 
                     ESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAH 
                     MIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSK 
                     SRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLD 
                     NLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTL 
                     LKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVK 
                     LNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPY 
                     YVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVL 
                     PKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKE 
                     DYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLT 
                     LFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDF 
                     LKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQT 
                     VKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKE 
                     HPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKV 
                     LTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKA 
                     GFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF 
                     YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIG 
                     KATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS 
                     MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVV 
                     AKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLF 
                     ELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQH 
                     KHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGA 
                     PAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDGSPKKKRKVS 
                     SDYKDHDGDYKDHDIDYKDDDDKAAG*"
     promoter        complement(3386..3490)
                     /label=AmpR promoter
                     /note="/Transferred_From=pCMV-T7-Cas9 (MLM3613), 
                     pCMV-T7-Cas9 delta NcoI"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=JDS246; 
                     pCMV-NLS-hTtAgo-NLS-3xFLAG; 
                     pLV-CMVEI.EGFPDS-iNpLTRSEBBA6_0693; pLVX-TetOne-puro; 
                     pMLM3705; pMLM3705 annotation mess"
                     /note="/X-Transferred_Similarity=100.00%"
     misc_feature    3774..4137
                     /label=CMV Enhancer
                     /note="/Transferred_From=pCMV-T7-Cas9 (MLM3613), 
                     pCMV-T7-Cas9 delta NcoI"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=3. 
                     pCAG-nls-Csy4-FokI-T2A-dCas9-recoded (C-terminal fusion) - 
                     linker variants; 6.pCAG-nls-FokI-Csy4-T2A-Cas9-recoded 
                     (N-terminal fusion) -linker variants; JDS246; 
                     pCAG-Csy4-P2A-NLS-FLAG-NLS-dCas9-NLS-FLAG-VP64 recoded 
                     (BPK376); pCAG-Csy4-P2A-NLS-FLAG-NLS-dCas9-NLS-FLAG-VP64 
                     recoded (BPK376frameshift); 
                     pCAG-Csy4-P2A-dCas9-NLS-FLAG-VP64 recoded; 
                     pCAG-Csy4-T2A-Cas9-VP64 recoded (BPK1); 
                     pCAG-Csy4-T2A-Cas9-VP64 recoded (BPK2a); 
                     pCAG-Csy4-T2A-Cas9-recoded (SQT834); 
                     pCAG-Csy4-T2A-FokI-dCas9-recoded-nls (JAF1484); 
                     pCAG-Csy4-T2A-KRAB-dCas9-recoded-nlsFLAG (NW9); 
                     pCAG-Csy4-T2A-NLS-FLAG-NLS-VP64-dCas9-recoded (making 
                     p125); pCAG-Csy4-T2A-NLS-FLAG-NLS-dCas9-KRAB recoded 
                     (BPK353); pCAG-Csy4-T2A-NLS-FLAG-NLS-dCas9-VP64 recoded 
                     (BPK326); pCAG-Csy4-T2A-NLS-FLAG-NLS-dCas9-VP64 recoded NO 
                     CTERM FLAG; 
                     pCAG-Csy4-T2A-VP64-NLS/FLAG/NLS-dCas9-recoded-nlsFLAG 
                     (BPK328); pCAG-Csy4-T2A-VP64-dCas9 linker variant BbsI 
                     cassette; pCAG-Csy4-T2A-VP64-dCas9-recoded-nlsFLAG 
                     (BPK285); pCAG-Csy4-T2A-VP64-dCas9-recoded-nlsFLAG 
                     (BPK286); pCAG-Csy4-T2A-dCas9-KRAB recoded (NW8); 
                     pCAG-Csy4-T2A-dCas9-VP64 recoded (BPK3); 
                     pCAG-Csy4-T2A-dCas9-VP64 recoded NO FLAG (BPK373 or 374?); 
                     pCAG-NLS-FLAG-NLS-dCas9-NLS-FLAG-VP64; 
                     pCAG-NLS-FLAG-NLS-dCas9-VP64 recoded; 
                     pCAG-NLS-FLAG-VP64-dCas9-recoded; 
                     pCAG-NLS-VP64-dCas9-recoded; pCAG-NLS-dCas9-NLS-3xFLAG 
                     recoded; pCAG-NLS-dCas9-VP64 recoded; pCAG-NLS-dCas9-VP64 
                     recoded CopyXXXXX; 
                     pCAG-NLS-hCsy4-P2A-NLS-FLAG-NLS-hdCas9-NLS-FLAG-VP64; 
                     pCAG-NLS-hTtAgo (BPK633); pCAG-NLS-hTtAgo-NLS-3xFLAG 
                     (BPK573); pCAG-NLS-hTtAgo-NLS-3xFLAG-VP64; 
                     pCAG-NLS-hTtAgo-NLS-3xFLAG-VP64 D478A/D546A; 
                     pCAG-NLS-hTtAgo-NLS-3xFLAG-VP64 D478N/D546N; 
                     pCAG-VP64-NLS-FLAG-NLS-dCas9-recoded-NLS-FLAG (BPK329); 
                     pCAG-VP64-NLS-hTtAgo; pCAG-VP64-dCas9-recod linker variant 
                     BbsI cassette; pCAG-VP64-dCas9-recoded GGGGSx5 linker; 
                     pCAG-VP64-dCas9-recoded-NLS-FLAG (BPK278); 
                     pCAG-VP64-hCsy4-NLS-P2A-NLS-hdCas9-NLS-FLAG-VP64; 
                     pCAG-dCas9-VP64; 
                     pCAG-hCsy4-NLS-P2A-NLS-FLAG-NLS-hdCas9-NLS-FLAG-VP64 
                     (BPK553); pCAG-hCsy4-NLS-VP64-P2A-NLS-hdCas9-NLS-FLAG-VP64;
                     pCAG-hCsy4-P2A-NLS-FLAG-NLS-dCas9-NLS-3xFLAG-KRAB (BPK550);
                     pCAG-hCsy4-P2A-NLS-FLAG-NLS-dCas9-NLS-FLAG-VP64 
                     delta175-307 (BPK544); 
                     pCAG-hCsy4-P2A-NLS-FLAG-NLS-dCas9-NLS-FLAG-VP64 recoded; 
                     pCAG-hCsy4-P2A-NLS-FLAG-NLS-dCas9-NLS-FLAG-VP64 recoded 
                     (BPK447); 
                     pCAG-hCsy4-P2A-NLS-FLAG-NLS-dCas9-NLS-FLAG-VP64-GG recoded 
                     (BPK531); pCAG-hCsy4-P2A-NLS-FLAG-NLS-dCas9-NLS-KRAB 
                     (BPK454); pCAG-hCsy4-P2A-NLS-FLAG-NLS-dCas9-NLS-VP64 
                     recoded (BPK453); pCAG-hCsy4-P2A-NLS-dCas9-NLS-FLAG-VP64 
                     recoded (BPK546); pCAG-hCsy4-P2A-NLS-dCas9-NLS-KRAB 
                     (BPK548); pCAG-hCsy4-P2A-dCas9-NLS-FLAG-VP64 recoded 
                     (BPK422); pCAG-hCsy4-P2A-dCas9-NLS-KRAB (BPK439); 
                     pCAG-hCsy4-P2A-dCas9-NLS-VP64 recoded (BPK450); 
                     pCAG-hCsy4-T2A-NLS-FLAG-NLS-dCas9-NLS-FLAG-VP64 recoded 
                     (BPK428); pCAG-hCsy4-T2A-NLS-FLAG-NLS-dCas9-NLS-KRAB 
                     (BPK444); pCAG-hCsy4-T2A-NLS-FLAG-NLS-dCas9-NLS-VP64 
                     recoded (BPK437); pCAG-hCsy4-T2A-dCas9-NLS-FLAG-VP64 
                     recoded (BPK426); pCAG-hCsy4-T2A-dCas9-NLS-KRAB (BPK442); 
                     pCAG-hCsy4-T2A-dCas9-NLS-VP64 recoded (BPK435); 
                     pCAG-hCsy4-T2A-nls-hFokI-dhCas9-nls-3XFLAG (SQT1601); 
                     pCAG-hTtAgo; pCAG-hTtAgo-NLS; pCAG-hTtAgo-NLS-3xFLAG; 
                     pCAG-nls-FokI-Csy4-T2A-dCas9-recoded (N-terminal fusion 
                     CK1891); pCMV-NLS-hTtAgo-NLS-3xFLAG; pLenti-CAG-Puro-DEST; 
                     pMLM3705; pMLM3705 annotation mess"
                     /note="/X-Transferred_Similarity=100.00%"
     promoter        complement(3823..3851)
                     /label=AmpR promoter
                     /note="/Transferred_From=pCMV-T7-Cas9 (MLM3613), 
                     pCMV-T7-Cas9 delta NcoI"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_Similarity=100.00%"
                     /note="/modified_by=User"
     misc_feature    3840..4127
                     /label=CAG enhancer
                     /note="/Transferred_From=pCMV-T7-Cas9 (MLM3613), 
                     pCMV-T7-Cas9 delta NcoI"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_Similarity=100.00%"
                     /note="/modified_by=User"
     promoter        4295..4364
                     /label=CMV promoter
                     /note="/modified_by=User"
     promoter        4385..4404
                     /label=T7 promoter
                     /note="/Transferred_From=pCMV-T7-Cas9 (MLM3613), 
                     pCMV-T7-Cas9 delta NcoI"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=pCRII-TOPO"
                     /note="/X-Transferred_Similarity=100.00%"
                     /note="/modified_by=User"
     misc_feature    join(4411..4416,1..4)
                     /label=Kozak sequence
                     /label=nonstandard type: motif
                     /note="/Transferred_From=pCMV-T7-Cas9 (MLM3613), 
                     pCMV-T7-Cas9 delta NcoI"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=JDS246; 
                     VC-(VC780+Csy4-Cas9)_pLKO-TRC001; 
                     pCAG-NLS-FLAG-NLS-dCas9-NLS-FLAG-VP64; 
                     pCAG-NLS-dCas9-NLS-3xFLAG recoded; pCAG-NLS-hTtAgo 
                     (BPK633); pCAG-NLS-hTtAgo-NLS-3xFLAG (BPK573); 
                     pCAG-NLS-hTtAgo-NLS-3xFLAG-VP64 D478A/D546A; 
                     pCAG-NLS-hTtAgo-NLS-3xFLAG-VP64 D478N/D546N; 
                     pCAG-VP64-NLS-hTtAgo; 
                     pCAG-hCsy4-NLS-VP64-P2A-NLS-hdCas9-NLS-FLAG-VP64; 
                     pCAG-hTtAgo; pCAG-hTtAgo-NLS; pCAG-hTtAgo-NLS-3xFLAG; 
                     pCW-Cas9 (modified); pCW-EFS-Puro-P2A-NFN-dCas9-VP64; 
                     pCW-NLS-dCas9-VP64 EFS-Puro-T2A-rTta3-WPRE; 
                     pLV-CMVEI.EGFPDS-iNpLTRSEBBA6_0693; 
                     pLenti-TREminCMV-NLS-3xFLAG-NLS-dCas9-VP64 
                     EFS-Puro-T2A-rTta3-WPRE; pLenti-TREminCMV-NLS-dCas9-VP64 
                     EFS-Puro-T2A-rTta3-WPRE; pMLM3705; pMLM3705 annotation 
                     mess; pWPXLd (modified)"
                     /note="/X-Transferred_Similarity=100.00%"
                     /note="/modified_by=User"
ORIGIN
        1 atgggccatc acaccatcgc ccgcatgaat gctttcaaca aggcttttgc aaacgctaaa
       61 gactgctaca aaaaaatgca agcctggcac ctgttgaaca aaccgaaaca cgcattcttt
      121 cctatgcaga acacgccagc cctggataac ggcctcgctg cgctttatga actccggggg
      181 ggaaaggaag atgcacatat tttgtctatt ctcagcagat tgtacttgta cggagcgtgg
      241 cggaatacat tgggcatata ccagttggac gaggagatca ttaaggactg caaggagctt
      301 cccgacgata caccgacgag catattcctc aacttgcctg actggtgtgt atatgtagat
      361 atatccagtg ctcagatcgc cactttcgac gacggcgtag cgaagcacat caaaggtttc
      421 tgggccatat acgatattgt tgaaatgaat ggaatcaacc atgacgtatt ggacttcgtt
      481 gtagatactg acaccgacga taatgtgtac gttccgcagc ctttcatttt gtcatctggt
      541 caatccgtgg cagaagtgct tgactatgga gcaagtttgt tcgacgacga tactagcaat
      601 acgcttatta agggcctcct gccgtacttg ctctggcttt gcgtagcaga acccgacatc
      661 acttacaaag gtttgccagt ctcaagagag gaacttactc gaccgaaaca ttcaatcaat
      721 aaaaaaacgg gcgcgtttgt gacacctagt gagccattca tctaccagat aggagaacgg
      781 ctgggaagtg aggttaggag gtatcagagc attatagatg gggagcaaaa gcgcaatcga
      841 ccacatacaa agcgccctca tattcgccgc ggtcactggc atggatattg gcaagggact
      901 ggacaggcga aggagtttcg agttcgatgg caacccgctg tgttcgtgaa tagcggaaga
      961 gtatcatccg gatccggcgg tggaggcagt gggcccaaga agaagaggaa agtctgaccg
     1021 gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta
     1081 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     1141 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     1201 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     1261 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     1321 gctcgatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     1381 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     1441 aagcctaggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     1501 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     1561 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     1621 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1681 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1741 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1801 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1861 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1921 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1981 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2041 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2101 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2161 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2221 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2281 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2341 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2401 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2461 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2521 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     2581 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     2641 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     2701 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     2761 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2821 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     2881 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2941 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     3001 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3061 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     3121 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     3181 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     3241 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     3301 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     3361 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     3421 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     3481 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcgat
     3541 ctcccgatcc cctagggtcg actctcagta caatctgctc tgatgccgca tagttaagcc
     3601 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     3661 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     3721 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
     3781 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     3841 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     3901 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     3961 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     4021 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     4081 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     4141 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     4201 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     4261 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     4321 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag agatccgcgg
     4381 ccgctaatac gactcactat agggagagcc gccacc
//
