LOCUS pPS 6593 bp DNA circular UNA 30-DEC-2016 DEFINITION Gibson Assembly of 1_gBlock_site-site-scaf-BsmBI-hU6 into 1_PmeI-HpaI_WeissmanPlasmid. ACCESSION urn.local...6vsu9by-y VERSION urn.local...6vsu9by-y KEYWORDS . SOURCE ORGANISM . FEATURES Location/Qualifiers origin 0^1 /label="origin" promoter 241..269 /ApEinfo_revcolor="#f58a5e" /ApEinfo_fwdcolor="#f58a5e" /label="AmpR promoter" promoter 595..1158 /ApEinfo_revcolor="#c6c9d1" /ApEinfo_fwdcolor="#c6c9d1" /label="pU6 5'CMV promoter" LTR 1176..1356 /ApEinfo_revcolor="#85dae9" /ApEinfo_fwdcolor="#85dae9" /label="LTR 5'" misc_feature 1408..1545 /created_by="User" /label="Psi" misc_feature 1512..1882 /created_by="User" /label="Gag" misc_feature 2028..2269 /created_by="User" /label="RRE" misc_feature 2787..2904 /created_by="User" /label="cPPT" primer_bind 2913..2932 /Sequence="TTACAGGGACAGCAGAGATC" /Hairpin_Tm="None" /Tm="56.3" /Self_Dimer_Tm="None" /%GC="50.0" /created_by="primer3" /label="pHR-F" polyA_signal complement(2963..3190) /ApEinfo_revcolor="#c7b0e3" /ApEinfo_fwdcolor="#c7b0e3" /modified_by="User" /label="ANTIPARALLEL - BGH pA" misc_feature 3184..3738 /Original_Bases /label /note="Geneious type: Editing History Insertion" ligation 3184..3208 /label="Ligation" source 3209..3224 /created_by="User" /modified_by="User" /label="WeissmanPlasmid" misc_binding 3224..3243 /created_by="User" /modified_by="User" /label="GBC_CloningSite" motif 3224..3229 /created_by="User" /label="EcoRI" motif 3238..3243 /created_by="User" /label="EcoRI" misc_feature 3239..3738 /label="1_gBlock_site-site-scaf-BsmBI-hU6" /note="Geneious type: Inserted sequence" misc_feature <3244..>3295 /label="1_gBlock_site-site-scaf-BsmBI-hU6" /note="Geneious type: Inserted sequence" source 3244..3248 /created_by="User" /label="WeissmanPlasmid" misc_binding 3272..3295 /created_by="User" /modified_by="User" /label="EF1a-Puro-BFP_CloningSite" motif 3272..3279 /created_by="User" /modified_by="User" /label="NotI" motif 3288..3295 /created_by="User" /modified_by="User" /label="NotI" misc_RNA complement(3319..3410) /ApEinfo_fwdcolor="#ff3d4c" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /modified_by="User" /label="Optimized scaffold sgRNA" misc_binding 3411..3463 /note="Filler" /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="pink" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /modified_by="User" /label="Filler" motif complement(3412..3417) /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /modified_by="User" /label="BsmBI" motif 3453..3458 /ApEinfo_label="BsmBI recognition site" /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /modified_by="User" /label="BsmBI" promoter complement(3460..3708) /note="hU6" /ApEinfo_fwdcolor="#c48d3c" /ApEinfo_revcolor="pink" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /label="hU6" misc_feature 3526..3531 /Recognition_pattern="CA^TATG" /annotation_group="CATATG" /label="NdeI, FauNDI" /note="Geneious type: restriction site" ligation 3709..3738 /label="Ligation" LTR 3804..4404 /ApEinfo_revcolor="#faac61" /ApEinfo_fwdcolor="#faac61" /modified_by="User" /label="WPRE" misc_feature complement(3867..3887) /Sequence="CATAGCGTAAAAGGAGCAACA" /Hairpin_Tm="None" /Tm="56.6" /Self_Dimer_Tm="None" /%GC="42.9" /created_by="primer3" /label="WPRE-R" /note="Geneious type: primer_bind_reverse" LTR 4594..4608 /ApEinfo_revcolor="#9eafd2" /ApEinfo_fwdcolor="#9eafd2" /label="cPPT" LTR 4931..5111 /ApEinfo_revcolor="#c6c9d1" /ApEinfo_fwdcolor="#c6c9d1" /label="3' LTR" rep_origin 5155..5837 /ApEinfo_revcolor="#b7e6d7" /ApEinfo_fwdcolor="#b7e6d7" /label="ColE1 origin" gene complement(5935..1) /ApEinfo_revcolor="#ff9ccd" /ApEinfo_fwdcolor="#ff9ccd" /label="AmpR" ORIGIN 1 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 61 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 121 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 181 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 241 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 301 cgcgcacatt tccccgaaaa gtgccacctg acgcttaagc ggtcgacgga tcgggagatc 361 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca 421 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc 481 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt 541 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta 601 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac 661 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc 721 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt 781 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac 841 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac 901 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt 961 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc 1021 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt 1081 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg 1141 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct 1201 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 1261 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 1321 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa 1381 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac 1441 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta 1501 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg 1561 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg 1621 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg 1681 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag 1741 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga 1801 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca 1861 gcaagcggcc ggccgcgctg atcttcagac ctggaggagg agatatgagg gacaattgga 1921 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca 1981 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc 2041 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac 2101 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg 2161 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa 2221 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg 2281 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg 2341 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa 2401 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat 2461 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt 2521 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg 2581 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc 2641 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga 2701 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc 2761 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt 2821 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa 2881 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg 2941 ttagtaccgg gcccgctcta gaccatagag cccaccgcat ccccagcatg cctgctattg 3001 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac 3061 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca 3121 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa 3181 ggcacagtcg aggctgatca gcgggtttaa acgggccctc taggaattca tgcatgcgaa 3241 ttcaattcgc cagggttttc ccagtcacga cgcggccgca tgcatgcgcg gccgccacca 3301 aacgtggctt gccagcccaa aaaagcaccg actcggtgcc actttttcaa gttgataacg 3361 gactagcctt atttaaactt gctatgctgt ttccagcata gctcttaaac agagacgttg 3421 aagatgaaac gtgtagtgtg ctcatttaca accgtctccg gtgtttcgtc ctttccacaa 3481 gatatataaa gccaagaaat cgaaatactt tcaagttacg gtaagcatat gatagtccat 3541 tttaaaacat aattttaaaa ctgcaaacta cccaagaaat tattactttc tacgtcacgt 3601 attttgtact aatatctttg tgtttacagt caaattaatt ctaattatct ctctaacagc 3661 cttgtatcgt atatgcaaat atgaaggaat catgggaaat aggccctcaa cgtcgaggga 3721 cctaataact tcgtatagca tacattatac gaagttatac atgtttaagg gttccggttc 3781 cactaggtac aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt 3841 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct 3901 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat 3961 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg 4021 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag 4081 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc 4141 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg 4201 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc 4261 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc 4321 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc 4381 tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc tagaaaaaca 4441 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc 4501 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac 4561 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 4621 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta 4681 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg 4741 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga 4801 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt 4861 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc 4921 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact 4981 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc 5041 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa 5101 aatctctagc agcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 5161 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 5221 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 5281 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 5341 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 5401 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 5461 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 5521 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 5581 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 5641 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 5701 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 5761 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 5821 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 5881 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 5941 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 6001 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 6061 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 6121 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 6181 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 6241 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 6301 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 6361 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 6421 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 6481 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 6541 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tca //