LOCUS Cellecta-pRCCR-C 11309 bp ds-DNA circular SYN 13-DEC-2018 DEFINITION CRISPR Cas9 pRCCR-CMV-Cas9-2A-TagRFP (plasmid). SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11309) AUTHORS Cellecta TITLE Direct Submission JOURNAL Exported Dec 13, 2018 from SnapGene 4.2.9 to Vector NTI(R) format http://www.snapgene.com COMMENT VNTDATE|728524800| COMMENT VNTDBDATE|860976000| COMMENT VNTNAME|Cellecta-pRCCR-CMV-Cas9-2A-TagRFP.dna| COMMENT VNTAUTHORNAME|Cellecta| FEATURES Location/Qualifiers promoter 6..232 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 233..413 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 460..585 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1078..1311 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." misc_feature 1818..1933 /label=cPPT /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" promoter 2005..2348 /label=CMV CDS 2369..2389 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS CDS 2393..2413 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS CDS 2417..2437 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS CDS 2444..6544 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" CDS 6545..6592 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS CDS 6608..6661 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." CDS 6668..7381 /codon_start=1 /product="monomeric derivative of red fluorescent protein from Entacmaea quadricolor (Merzlyak et al., 2007)" /label=TagRFP /note="mammalian codon-optimized" misc_feature 7388..7976 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 8050..8283 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 8355..8476 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 8516..8651 /label=SV40 ori /note="SV40 origin of replication" rep_origin complement(9086..9674) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(9845..10705) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" promoter complement(10706..10810) /gene="bla" /label=AmpR promoter ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct 241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc 421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg 481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt 541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg 601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa 661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg 721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca 781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa 841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa 901 agtaagacca ccgcacagca agcggccact gatcttcaga cctggaggag gagatatgag 961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt 1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg 1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac 1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct 1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct 1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg 1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa 1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa 1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa 1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac 1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag 1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc 1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga 1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg 1801 ttgcttggat cgagcaattt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa 1861 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa 1921 aaattcaaaa tttctgcgtt gttgtcggtg ctcgttctct gctcttcacg ctactgaatt 1981 catcaccggt tcttcgaatc tagtattatg cccagtacat gaccttatgg gactttccta 2041 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 2101 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 2161 acgccaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 2221 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtggaaggtt tatataagca 2281 gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc 2341 atagaagatt ctagccgcca ccatggcccc aaaaaagaag agaaaggtag atccaaaaaa 2401 gaagagaaag gtagatccaa aaaagaagag aaaggtaggt ggagacaaga agtacagcat 2461 cggcctggac atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt 2521 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct 2581 gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac 2641 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag 2701 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt 2761 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc 2821 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga 2881 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca 2941 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca 3001 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga 3061 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc 3121 ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg 3181 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag 3241 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc 3301 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag 3361 agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga 3421 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta 3481 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc 3541 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga 3601 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa 3661 cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga 3721 agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg 3781 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag 3841 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc 3901 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt 3961 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt 4021 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc 4081 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga 4141 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt 4201 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct 4261 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga 4321 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt 4381 gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat 4441 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg 4501 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga 4561 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct 4621 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct 4681 cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa 4741 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg 4801 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca 4861 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga 4921 actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct 4981 gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag 5041 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct 5101 gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg 5161 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat 5221 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga 5281 caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg 5341 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc 5401 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga 5461 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca 5521 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa 5581 gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg 5641 cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct 5701 gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa 5761 agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga 5821 ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc 5881 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac 5941 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta 6001 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga 6061 aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc 6121 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg 6181 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga 6241 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct 6301 ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga 6361 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt 6421 tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct 6481 gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg 6541 cgacaaaagg ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa aggcggccgc 6601 aggatccgag ggcagaggaa gccttctaac atgcggtgac gtggaggaga atcccggccc 6661 ttccggaatg gtgtctaagg gcgaagagct gattaaggag aacatgcaca tgaagctgta 6721 catggagggc accgtgaaca accaccactt caagtgcaca tccgagggcg aaggcaagcc 6781 ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc 6841 cttcgacatc ctggctacca gcttcatgta cggcagcaga accttcatca accacaccca 6901 gggcatcccc gacttcttta agcagtcctt ccctgagggc ttcacatggg agagagtcac 6961 cacatacgag gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg 7021 cctcatctac aacgtcaaga tcagaggggt gaacttccca tccaacggcc ctgtgatgca 7081 gaagaaaaca ctcggctggg aggccaacac cgagatgctg taccccgctg acggcggcct 7141 ggaaggcaga agcgacatgg ccctgaagct cgtgggcggg ggccacctga tctgcaactt 7201 caagaccaca tacagatcca agaaacccgc taagaacctc aagatgcccg gcgtctacta 7261 tgtggaccac agactggaaa gaatcaagga ggccgacaaa gagacctacg tcgagcagca 7321 cgaggtggct gtggccagat actgcgacct ccctagcaaa ctggggcaca aacttaattg 7381 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 7441 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 7501 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 7561 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 7621 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 7681 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 7741 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg 7801 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 7861 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 7921 gcgtctccgc cttcgccctc agacgagtcg gatctctctt tgggccgcct ccccgcctgg 7981 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa 8041 aggggggact ggaagggcta attcactccc aacgaagata agatctgctt tttgcttgta 8101 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 8161 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 8221 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 8281 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat 8341 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 8401 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 8461 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc 8521 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 8581 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt 8641 ttttggaggc ctagactttt gcagagacca aattcgtaat catgtcatag ctgtttcctg 8701 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 8761 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 8821 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 8881 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 8941 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 9001 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 9061 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 9121 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 9181 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 9241 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 9301 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 9361 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 9421 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 9481 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 9541 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 9601 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 9661 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 9721 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 9781 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 9841 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 9901 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 9961 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 10021 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 10081 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 10141 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 10201 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 10261 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 10321 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 10381 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 10441 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 10501 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 10561 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 10621 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 10681 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 10741 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 10801 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca 10861 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg 10921 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag 10981 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg 11041 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg 11101 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc 11161 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga 11221 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac 11281 gttgtaaaac gacggccagt gccaagctg //