LOCUS Exported 8507 bp ds-DNA circular SYN 15-DEC-2016 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pY036_ATP1A1_G5_Array SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8507) AUTHORS . TITLE Direct Submission JOURNAL Exported Thursday, Dec 15, 2016 from SnapGene 3.2.1 http://www.snapgene.com COMMENT Ligation of : pY036 U6(Bbs1)-CBh-NLS-AsCpf1-NLS-3xHA-pA [808 9 nt] : (Acc65I[520] / XbaI[148]) to BbsI_DR_APT1A1G5_Gbloc k_2 [410 nt] : (XbaI[7] / Acc65I[421]) FEATURES Location/Qualifiers source 1..8507 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 1..141 /note="L-ITR" /note="dnas_title=""L-ITR"" - vntifkey=""21"" - label=L-ITR - 1..141" misc_feature 148..153 /note="XbaI_misc_feature" /note="note=""XbaI"" - 148..153" misc_feature 154..483 /note="KpnI_misc_feature [5']" /note="note=""KpnI"" - 154..525" promoter 154..402 /note="Human U6 Promoter" /note="dnas_title=""Human U6 Promoter"" - vntifkey=""29"" - label=Human U6 Promoter - 154..402" misc_feature 154..402 /note="Human U6 promoter" /note="note=""direct strand translation""translation=""EGLFPMIPSYLHIRYKAVREIIGINLTVNTKI LVQNT*""" misc_feature 154..178 /note="PCR_primer" /note="pair="" - primer=""GAGGGCCTATTTCCCATGATTCCTT"" - current=0 - 154..178" misc_feature complement(372..402) /note="PCR_primer" /note="pair="" - primer=""GGTGTTTCGTCCTTTCCACAAGATATATAAA"" - current=0 - complement(372..402)" misc_feature 403 /note="G" /note="note=""direct strand translation""translation=""X""" misc_feature 404..423 /note="5'DR 20nt_misc_feature" /note="note=""5'DR 20nt"" - 404..423" misc_feature 404..423 /note="DR #1 (20bp)" /note="note=""direct strand translation""translation=""*""" misc_feature complement(415..447) /note="PCR_primer" /note="pair="" - primer=""AAAAAGAGACGATATATCGTCTCCATCTACAAG"" - current=0 - complement(415..447)" misc_feature 420..423 /note="Bbs1_misc_feature" /note="note=""Bbs1"" - 420..423" misc_feature 426..431 /note="BbsI" /note="note=""direct strand translation""translation=""VF""" misc_feature 434..439 /note="BbsI" /note="note=""direct strand translation""translation=""ED""" misc_feature 441..460 /note="5'DR 20nt_misc_feature" /note="note=""5'DR 20nt"" - 404..423" misc_feature 442..460 /note="DR #2 (19bp)" misc_feature 442..445 /note="Bbs1_misc_feature" /note="note=""Bbs1"" - 442..445" misc_feature 461..483 /note="ATP1A1_G5 (23bp)" misc_feature 484..489 /note="polIII terminator" misc_feature 526..567 /note="KpnI_misc_feature [3']" /note="note=""KpnI"" - 154..525" promoter 561 /note="CMV promoter" /note="dnas_title=""CMV promoter"" - invitrogen=""2180000"" - vntifkey=""29"" - label=CMV promoter - 519..519" misc_feature 568..1384 /note="CBh_misc_feature" /note="note=""CBh"" - 526..1342" misc_feature 1385 /note="AgeI_misc_feature" /note="note=""AgeI"" - 1343..1343" misc_feature 1406..1409 /note="Golden Gate_misc_feature" /note="note=""Golden Gate"" - 1364..1367" misc_feature 1417..5523 /note="AsCpf1_2x_NLS_3x_HA" /note="note=""direct strand translation""translation=""MAPKKKRKVGIHGVPAATQFEGFTNLYQVSKT LRFELIPQGKTLKHIQEQGFIEEDKARNDHYKELKPIIDRIYKTYADQCLQLVQLDWEN LSAAIDSYRKEKTEETRNALIEEQATYRNAIHDYFIGRTDNLTDAINKRHAEIYKGLFK AELFNGKVLKQLGTVTTTEHENALLRSFDKFTTYFSGFYENRKNVFSAEDISTAIPHRI " misc_feature 1417..1467 /note="NLS-SV40_misc_feature" /note="note=""NLS-SV40"" - 1375..1425" misc_feature 1468..5385 /note="huAsCpf1_misc_feature" /note="note=""huAsCpf1"" - 1426..5343" misc_feature complement(4163..4185) /note="PCR_primer" /note="pair="" - primer=""GATGCCGATGATAGGTGTCTCGG"" - current=0 - complement(4121..4143)" misc_feature 4174..4665 /note="RuvC_misc_feature" /note="note=""RuvC"" - 4132..4623" misc_feature 4186..4188 /note="D908A_misc_feature" /note="note=""D908A"" - 4144..4146" misc_feature 4190..4216 /note="PCR_primer" /note="pair="" - primer=""GGGGCGAGAGAAACCTGATCTATATCA"" - current=0 - 4148..4174" misc_feature complement(4422..4440) /note="PCR_primer" /note="pair="" - primer=""CAGCACCACCACGGCCTGG"" - current=0 - complement(4380..4398)" misc_feature 4441..4443 /note="E993A_misc_feature" /note="note=""E993A"" - 4399..4401" misc_feature 4445..4470 /note="PCR_primer" /note="pair="" - primer=""ACCTGAATTTCGGCTTTAAGAGCAAG"" - current=0 - 4403..4428" misc_feature complement(4486..4509) /note="PCR_primer" /note="pair="" - primer=""GAACTGCTGGTACACGGCCTTCTC"" - current=0 - complement(4444..4467)" misc_feature 4510..4512 /note="E1016A_misc_feature" /note="note=""E1016A"" - 4468..4470" misc_feature 4514..4539 /note="PCR_primer" /note="pair="" - primer=""AGATGCTGATCGATAAGCTGAATTGC"" - current=0 - 4472..4497" misc_feature 4672..4956 /note="Zinc Finger_misc_feature" /note="note=""Zinc Finger"" - 4630..4914" misc_feature 5107..5244 /note="RuvC III_misc_feature" /note="note=""RuvC III"" - 5065..5202" misc_feature complement(5149..5166) /note="PCR_primer" /note="pair="" - primer=""CTCGCCTGTGGCGGCATT"" - current=0 - complement(5107..5124)" misc_feature 5167..5169 /note="D1235A_misc_feature" /note="note=""D1235A"" - 5125..5127" misc_feature 5171..5190 /note="PCR_primer" /note="pair="" - primer=""ATATCAACAGCCCCGTGCGC"" - current=0 - 5129..5148" misc_feature complement(5232..5250) /note="PCR_primer" /note="pair="" - primer=""GGCGTCCATGGGCCACTCT"" - current=0 - complement(5190..5208)" misc_feature complement(5239..5256) /note="PCR_primer" /note="pair="" - primer=""GGCATCGGCGTCCATGGG"" - current=0 - complement(5197..5214)" misc_feature 5251..5253 /note="D1263A_misc_feature" /note="note=""D1263A"" - 5209..5211" misc_feature 5255..5275 /note="PCR_primer" /note="pair="" - primer=""CCAATGGCGCCTACCACATCG"" - current=0 - 5213..5233" misc_feature 5257..5259 /note="N1265A_misc_feature" /note="note=""N1265A"" - 5215..5217" misc_feature 5261..5279 /note="PCR_primer" /note="pair="" - primer=""GCGCCTACCACATCGCCCT"" - current=0 - 5219..5237" misc_feature 5386..5433 /note="NLS nuc_misc_feature" /note="note=""NLS nuc"" - 5344..5391" misc_feature 5386..5433 /note="Nucleoplasmin NLS" /note="note=""direct strand translation""translation=""KRPAATKKAGQAKKKK""" misc_feature 5434..5439 /note="GS_misc_feature" /note="note=""GS"" - 5392..5397" misc_feature complement(5440..5520) /note="3xHA_misc_feature" /note="note=""3xHA"" - complement(5398..5478)" misc_feature 5530..5761 /note="bGH polyA_misc_feature" /note="note=""bGH polyA"" - 5488..5719" misc_feature 5770..5910 /note="R-ITR" /note="dnas_title=""R-ITR"" - vntifkey=""21"" - label=R-ITR - 5728..5868" rep_origin 6002..6308 /direction=RIGHT /note="f1 Origin" /note="dnas_title=""f1 Origin"" - vntifkey=""33"" - label=f1 Origin - 5960..6266" misc_feature 6827..7684 /note="AmpR" /note="dnas_title=""AmpR"" - vntifkey=""21"" - label=AmpR - 6785..7642" misc_feature 7542..7547 /note="BsaI_misc_feature" /note="note=""BsaI"" - 7500..7505" rep_origin 7835..8502 /direction=RIGHT /note="pUC ori" /note="dnas_title=""pUC ori"" - vntifkey=""33"" - label=pUC ori - 7793..8460" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggcctct agagagggcc tatttcccat gattccttca 181 tatttgcata tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac 241 acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca 301 gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc 361 gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccgtaatttc tactcttgta 421 gatgggtctt cgagaagacc taatttctac tcttgtagat tagtacacat cagatatctt 481 ctcttttttc aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga 541 tgtacgggcc agatatacgc gggtacccgt tacataactt acggtaaatg gcccgcctgg 601 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac 661 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt 721 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa 781 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta 841 catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac 901 tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt 961 ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg 1021 cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct 1081 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc 1141 gcggcgggcg ggagtcgctg cgacgctgcc ttcgccccgt gccccgctcc gccgccgcct 1201 cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg 1261 gcccttctcc tccgggctgt aattagctga gcaagaggta agggtttaag ggatggttgg 1321 ttggtggggt attaatgttt aattacctgg agcacctgcc tgaaatcact ttttttcagg 1381 ttggaccggc tagcgtttaa acttaagctt gccaccatgg ccccaaagaa gaagcggaag 1441 gtcggtatcc acggagtccc agcagccaca cagttcgagg gctttaccaa cctgtatcag 1501 gtgagcaaga cactgcggtt tgagctgatc ccacagggca agaccctgaa gcacatccag 1561 gagcagggct tcatcgagga ggacaaggcc cgcaatgatc actacaagga gctgaagccc 1621 atcatcgatc ggatctacaa gacctatgcc gaccagtgcc tgcagctggt gcagctggat 1681 tgggagaacc tgagcgccgc catcgactcc tatagaaagg agaaaaccga ggagacaagg 1741 aacgccctga tcgaggagca ggccacatat cgcaatgcca tccacgacta cttcatcggc 1801 cggacagaca acctgaccga tgccatcaat aagagacacg ccgagatcta caagggcctg 1861 ttcaaggccg agctgtttaa tggcaaggtg ctgaagcagc tgggcaccgt gaccacaacc 1921 gagcacgaga acgccctgct gcggagcttc gacaagttta caacctactt ctccggcttt 1981 tatgagaaca ggaagaacgt gttcagcgcc gaggatatca gcacagccat cccacaccgc 2041 atcgtgcagg acaacttccc caagtttaag gagaattgtc acatcttcac acgcctgatc 2101 accgccgtgc ccagcctgcg ggagcacttt gagaacgtga agaaggccat cggcatcttc 2161 gtgagcacct ccatcgagga ggtgttttcc ttcccttttt ataaccagct gctgacacag 2221 acccagatcg acctgtataa ccagctgctg ggaggaatct ctcgggaggc aggcaccgag 2281 aagatcaagg gcctgaacga ggtgctgaat ctggccatcc agaagaatga tgagacagcc 2341 cacatcatcg cctccctgcc acacagattc atccccctgt ttaagcagat cctgtccgat 2401 aggaacaccc tgtctttcat cctggaggag tttaagagcg acgaggaagt gatccagtcc 2461 ttctgcaagt acaagacact gctgagaaac gagaacgtgc tggagacagc cgaggccctg 2521 tttaacgagc tgaacagcat cgacctgaca cacatcttca tcagccacaa gaagctggag 2581 acaatcagca gcgccctgtg cgaccactgg gatacactga ggaatgccct gtatgagcgg 2641 agaatctccg agctgacagg caagatcacc aagtctgcca aggagaaggt gcagcgcagc 2701 ctgaagcacg aggatatcaa cctgcaggag atcatctctg ccgcaggcaa ggagctgagc 2761 gaggccttca agcagaaaac cagcgagatc ctgtcccacg cacacgccgc cctggatcag 2821 ccactgccta caaccctgaa gaagcaggag gagaaggaga tcctgaagtc tcagctggac 2881 agcctgctgg gcctgtacca cctgctggac tggtttgccg tggatgagtc caacgaggtg 2941 gaccccgagt tctctgcccg gctgaccggc atcaagctgg agatggagcc ttctctgagc 3001 ttctacaaca aggccagaaa ttatgccacc aagaagccct actccgtgga gaagttcaag 3061 ctgaactttc agatgcctac actggcctct ggctgggacg tgaataagga gaagaacaat 3121 ggcgccatcc tgtttgtgaa gaacggcctg tactatctgg gcatcatgcc aaagcagaag 3181 ggcaggtata aggccctgag cttcgagccc acagagaaaa ccagcgaggg ctttgataag 3241 atgtactatg actacttccc tgatgccgcc aagatgatcc caaagtgcag cacccagctg 3301 aaggccgtga cagcccactt tcagacccac acaaccccca tcctgctgtc caacaatttc 3361 atcgagcctc tggagatcac aaaggagatc tacgacctga acaatcctga gaaggagcca 3421 aagaagtttc agacagccta cgccaagaaa accggcgacc agaagggcta cagagaggcc 3481 ctgtgcaagt ggatcgactt cacaagggat tttctgtcca agtataccaa gacaacctct 3541 atcgatctgt ctagcctgcg gccatcctct cagtataagg acctgggcga gtactatgcc 3601 gagctgaatc ccctgctgta ccacatcagc ttccagagaa tcgccgagaa ggagatcatg 3661 gatgccgtgg agacaggcaa gctgtacctg ttccagatct ataacaagga ctttgccaag 3721 ggccaccacg gcaagcctaa tctgcacaca ctgtattgga ccggcctgtt ttctccagag 3781 aacctggcca agacaagcat caagctgaat ggccaggccg agctgttcta ccgccctaag 3841 tccaggatga agaggatggc acaccggctg ggagagaaga tgctgaacaa gaagctgaag 3901 gatcagaaaa ccccaatccc cgacaccctg taccaggagc tgtacgacta tgtgaatcac 3961 agactgtccc acgacctgtc tgatgaggcc agggccctgc tgcccaacgt gatcaccaag 4021 gaggtgtctc acgagatcat caaggatagg cgctttacca gcgacaagtt ctttttccac 4081 gtgcctatca cactgaacta tcaggccgcc aattccccat ctaagttcaa ccagagggtg 4141 aatgcctacc tgaaggagca ccccgagaca cctatcatcg gcatcgatcg gggcgagaga 4201 aacctgatct atatcacagt gatcgactcc accggcaaga tcctggagca gcggagcctg 4261 aacaccatcc agcagtttga ttaccagaag aagctggaca acagggagaa ggagagggtg 4321 gcagcaaggc aggcctggtc tgtggtgggc acaatcaagg atctgaagca gggctatctg 4381 agccaggtca tccacgagat cgtggacctg atgatccact accaggccgt ggtggtgctg 4441 gagaacctga atttcggctt taagagcaag aggaccggca tcgccgagaa ggccgtgtac 4501 cagcagttcg agaagatgct gatcgataag ctgaattgcc tggtgctgaa ggactatcca 4561 gcagagaaag tgggaggcgt gctgaaccca taccagctga cagaccagtt cacctccttt 4621 gccaagatgg gcacccagtc tggcttcctg ttttacgtgc ctgccccata tacatctaag 4681 atcgatcccc tgaccggctt cgtggacccc ttcgtgtgga aaaccatcaa gaatcacgag 4741 agccgcaagc acttcctgga gggcttcgac tttctgcact acgacgtgaa aaccggcgac 4801 ttcatcctgc actttaagat gaacagaaat ctgtccttcc agaggggcct gcccggcttt 4861 atgcctgcat gggatatcgt gttcgagaag aacgagacac agtttgacgc caagggcacc 4921 cctttcatcg ccggcaagag aatcgtgcca gtgatcgaga atcacagatt caccggcaga 4981 taccgggacc tgtatcctgc caacgagctg atcgccctgc tggaggagaa gggcatcgtg 5041 ttcagggatg gctccaacat cctgccaaag ctgctggaga atgacgattc tcacgccatc 5101 gacaccatgg tggccctgat ccgcagcgtg ctgcagatgc ggaactccaa tgccgccaca 5161 ggcgaggact atatcaacag ccccgtgcgc gatctgaatg gcgtgtgctt cgactcccgg 5221 tttcagaacc cagagtggcc catggacgcc gatgccaatg gcgcctacca catcgccctg 5281 aagggccagc tgctgctgaa tcacctgaag gagagcaagg atctgaagct gcagaacggc 5341 atctccaatc aggactggct ggcctacatc caggagctgc gcaacaaaag gccggcggcc 5401 acgaaaaagg ccggccaggc aaaaaagaaa aagggatcct acccatacga tgttccagat 5461 tacgcttatc cctacgacgt gcctgattat gcatacccat atgatgtccc cgactatgcc 5521 taagaattcc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg 5581 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt 5641 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg 5701 gtggggtggg gcaggacagc aagggggagg attgggaaga gaatagcagg catgctgggg 5761 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc 5821 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag 5881 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc 5941 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg 6001 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 6061 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 6121 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 6181 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac 6241 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 6301 tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat 6361 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa 6421 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata 6481 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct 6541 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt 6601 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata 6661 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt 6721 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag 6781 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca 6841 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc 6901 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat 6961 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc 7021 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg 7081 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc 7141 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat 7201 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga 7261 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc 7321 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc 7381 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt 7441 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc 7501 tggctggttt attgctgata aatctggagc cggtgagcgt ggaagccgcg gtatcattgc 7561 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca 7621 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca 7681 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt 7741 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta 7801 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg 7861 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc 7921 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag 7981 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa 8041 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc 8101 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc 8161 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta 8221 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag 8281 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct 8341 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga 8401 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc 8461 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt //