LOCUS       Exported                3639 bp ds-DNA     circular SYN 29-AUG-2018
DEFINITION  CMV-T7 promoter expression plasmid for human codon optimized Orf2mor
            anti-CRISPR protein with C-terminal NLS  .
ACCESSION   urn.local...h-9a99gh3
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3639)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 29, 2018 from SnapGene 4.2.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3639
                     /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..192
                     /label=human codon optimized Orf2mor
                     /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             193..216
                     /label=GS linker
                     /note="/Color=#ff7400"
                     /note="/created_by=User"
                     /note="/modified_by=User"
     CDS             217..237
                     /label=NLS(SV40)
                     /note="/Color=#6bba70"
                     /note="/created_by=User"
     terminator      289..516
                     /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(605..632)
                     /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(642..665)
                     /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(685..697)
                     /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      775..906
                     /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(974..1593)
                     /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(1748..2608)
                     /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(2609..2713)
                     /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    2997..3360
                     /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(3046..3074)
                     /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    3063..3350
                     /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        3518..3587
                     /label=CMV promoter
                     /note="/modified_by=User"
     promoter        3608..3627
                     /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(3634..3639,1..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 atgggcaata acccgaaaac cccagaatac acgcgcaagg ctattagggc atacgagaag
       61 aatttggtga gaaagagcgt tacctttgat gtccgcaagg acgatgatat ggaactgctt
      121 aaaatgatcg aacaagacgg tcgcacattt gctcagatag cacgcaccgc ccttttggaa
      181 caccttcaaa aaggatccgg cggtggaggc agtgggccca agaagaagag gaaagtctga
      241 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt
      301 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
      361 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
      421 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
      481 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
      541 ggggctcgat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
      601 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
      661 gtaaagccta gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
      721 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
      781 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
      841 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      901 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      961 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1021 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1081 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1141 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1201 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1261 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1321 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1381 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1441 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1501 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1561 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1621 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1681 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1741 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1801 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     1861 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     1921 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     1981 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2041 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2101 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2161 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     2221 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2281 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     2341 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     2401 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     2461 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     2521 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     2581 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2641 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2701 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     2761 gatctcccga tcccctaggg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
     2821 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     2881 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     2941 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
     3001 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     3061 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     3121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     3181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     3241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     3301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     3361 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     3421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     3481 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     3541 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagagatccg
     3601 cggccgctaa tacgactcac tatagggaga gccgccacc
//
