LOCUS Construct_29 10694 bp DNA circular UNA 20-APR-2016 DEFINITION 1xTetO_H1p_20nt3_stgRNA. ACCESSION urn.local...1445897695822.44 (modified) VERSION urn.local...1445897695822.44 KEYWORDS . SOURCE Human ORGANISM Human . FEATURES Location/Qualifiers promoter 239..815 /created_by="User" /modified_by="User" /label="CMV_imm_early promoter" misc_feature 318..605 /created_by="User" /label="CAG enhancer" LTR 835..1015 /created_by="User" /modified_by="User" /label="HIV-1_5_LTR" misc_feature 1126..1170 /created_by="User" /modified_by="User" /label="HIV-1_psi_pack" primer_bind 1510..1555 /created_by="User" /label="HP13" misc_feature 1680..1913 /created_by="User" /label="RRE" misc_feature 2444..2459 /created_by="User" /label="cPPT" CDS complement(<2594..2598) /gene="AMP" /SECDrawAs="Gene" /SECStyleId=1 /label="AMP CDS" primer_bind 2594..2616 /created_by="primer3" /%GC="69.6" /Tm="64.72" /Hairpin="4.0" /Primer-Dimer="4.0" /Monovalent_Salt_Concentration="50.0" /Divalent_Salt_Concentration="0.0" /DNTP_Concentration="0.0" /Sequence="GGGGTTCCGCGCACATTTCCCCG" /Mismatches=0 /annotation_group="PrimerPlugin Group3c68cbfa-121b-4872-a7ab-08a32877e25e_urn:local:.:1365 712513487.384" /label="JP313" promoter 2630..2861 /created_by="User" /label="H1 Promoter" primer_bind complement(2788..2847) /created_by="User" /label="HP14" primer_bind 2828..2887 /created_by="User" /label="HP20" misc_feature 2843..2861 /created_by="User" /modified_by="User" /label="1x TetO" misc_feature 2866 /created_by="User" /label="+1 TSS" /note="Geneious type: transcription_start" gRNA 2867..2885 /created_by="User" /modified_by="User" /label="20nt-3" misc_feature 2886..2961 /Original_Bases /modified_by="User" /modified_by="User" /label="stgRNA scaffold" modified_base 2887..2888 /created_by="User" /label="TT->GG" modified_base 2913..2914 /created_by="User" /label="AA->CC" primer_bind complement(2944..2992) /created_by="User" /label="HP16" misc_signal 2962..2967 /Original_Bases /modified_by="User" /label="Terminator" primer_bind complement(2993..3012) /created_by="User" /label="JP1864-T7E1_seq_r" CDS 3012..3527 /gene="PGKp" /SECDrawAs="Gene" /SECStyleId=1 /label="PGKp CDS" misc_signal order(3538..3546,3538..3546) /created_by="User" /label="KOZAK" CDS 3547..4260 /gene="EBFP2" /SECDrawAs="Gene" /SECStyleId=1 /modified_by="User" /label="EBFP2 CDS" sig_peptide 4261..4326 /created_by="User" /modified_by="User" /label="p2a" CDS 4327..5358 /created_by="User" /label="hygroR" misc_feature 5388..5975 /created_by="User" /label="WPRE" misc_feature 6162..6183 /created_by="User" /modified_by="User" /label="U3PPt" misc_feature 6162..6177 /created_by="User" /label="cPPT" LTR 6499..6679 /created_by="User" /label="truncHIV-1_3_LTR" terminator 6708..6935 /created_by="User" /label="bGH_PA_terminator" rep_origin 6998..7304 /created_by="User" /label="f1_origin" misc_feature complement(7424..7639) /created_by="User" /label="SV40 enhancer" rep_origin 7603..7680 /created_by="User" /label="SV40 origin" promoter 7798..7865 /created_by="User" /label="EM7 promoter" gene 7866..8237 /created_by="User" /modified_by="User" /label="bleo" terminator 8373..8492 /created_by="User" /label="SV40_PA_terminator" promoter complement(8589..8618) /created_by="User" /modified_by="User" /label="lac_promoter" CDS complement(9701..10561) /created_by="User" /label="AmpR" promoter complement(10603..10630) /created_by="User" /label="AmpR promoter" ORIGIN 1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg 61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac 241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct 841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg 1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga 1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata 1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg 1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg 1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag 1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt 1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt 1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt 2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag 2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata 2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta 2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta 2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa 2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt 2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat 2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa 2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag 2581 agatccagtt tggggggttc cgcgcacatt tccccgaaaa gtgccacctg aatcctatgc 2641 ttcgaacgct gacgtcatca acccgctcca aggaatcgcg ggcccagtgt cactaggcgg 2701 gaacacccag cgcgcgtgcg ccctggcagg aagatggctg tgagggacag gggagtggcg 2761 ccctgcaata tttgcatgtc gctatgtgtt ctgggaaatc accataaacg tgaaatgtct 2821 ttggatttgg gaatcttata agtccctatc agtgatagag atcccaagtc gcgtgtagcg 2881 aagcagggtt agagctagaa atagcaagtt aacctaaggc tagtccgtta tcaacttgaa 2941 aaagtggcac cgagtcggtg cttttttcta gacccagctt tcttgtacaa agttggcatt 3001 agacgtcgag gaattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg 3061 cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca 3121 ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc 3181 ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac 3241 gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca 3301 atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg 3361 ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg 3421 ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc 3481 tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcatcc caagcttgcc 3541 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg 3601 gacggcgacg taaacggcca caagttcagc gtgaggggcg agggcgaggg cgatgccacc 3661 aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc 3721 accctcgtga ccaccctgag ccacggcgtg cagtgcttcg cccgctaccc cgaccacatg 3781 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc 3841 ttcttcaagg acgacggcac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc 3901 ctagtgaacc gcatcgagct gaagggcgtc gacttcaagg aggacggcaa catcctgggg 3961 cacaagctgg agtacaactt caacagccac aacatctata tcatggccgt caagcagaag 4021 aacggcatca aggtgaactt caagatccgc cacaacgtgg aggacggcag cgtgcagctc 4081 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacagc 4141 cactacctga gcacccagtc cgtgctgagc aaagacccca acgagaagcg cgatcacatg 4201 gtcctgctgg agttccgcac cgccgccggg atcactctcg gcatggacga gctgtacaag 4261 ggaagcggag ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct 4321 ggacctaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc 4381 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc 4441 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa 4501 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac 4561 attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg 4621 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg 4681 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa 4741 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg 4801 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat 4861 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc 4921 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag 4981 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg 5041 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg 5101 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt 5161 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc 5221 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat 5281 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccggatcgg 5341 gagatggggg aggctaacta agaattcgat atcaagctta tcgataatca acctctggat 5401 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5461 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 5521 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 5581 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 5641 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 5701 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 5761 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 5821 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 5881 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 5941 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgagaccta 6001 gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg 6061 ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga 6121 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg 6181 gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca 6241 caaggctact tccctgattg gcagaactac acaccagggc cagggatcag atatccactg 6301 acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat 6361 gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag 6421 agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag 6481 ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct 6541 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta 6601 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca 6661 gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc 6721 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc 6781 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg 6841 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa 6901 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg 6961 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 7021 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 7081 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 7141 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 7201 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 7261 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 7321 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga 7381 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt 7441 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 7501 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 7561 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 7621 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 7681 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 7741 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg 7801 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact 7861 aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag 7921 cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg 7981 ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc 8041 cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt 8101 cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg 8161 agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg 8221 tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg 8281 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 8341 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca 8401 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 8461 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 8521 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 8581 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 8641 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 8701 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 8761 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 8821 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 8881 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 8941 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 9001 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 9061 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 9121 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 9181 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 9241 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 9301 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 9361 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 9421 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 9481 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 9541 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 9601 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 9661 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 9721 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 9781 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 9841 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 9901 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 9961 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 10021 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 10081 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 10141 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 10201 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 10261 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 10321 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 10381 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 10441 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 10501 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 10561 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 10621 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 10681 aagtgccacc tgac //