LOCUS Exported 2936 bp ds-DNA circular SYN 24-AUG-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 2936) AUTHORS Bikard Lab TITLE Direct Submission JOURNAL Exported Aug 29, 2017 from SnapGene 4.0.3 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2936 /organism="recombinant plasmid" /mol_type="other DNA" primer_bind 1..18 /label=LC42 /note="ELZ2MutBB_R" primer_bind 73..92 /label=LC13 /note="pZS24-BB-R" primer_bind complement(94..116) /label=B377 pZ-F /note="from pDB129 pZE21-CRISPR-bsaI" primer_bind complement(103..125) /label=B385 CDS 209..1003 /codon_start=1 /label=kanR /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" misc_feature 1002..1031 /label=CBB CRISPR spapcer /note="cut pELZ2mut1K backbone, cut pCRISPR, pZE21, pCRRNA backbone " terminator 1034..1128 /label=lambda t0 terminator /note="transcription terminator from phage lambda" rep_origin 1216..1804 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1285..1304) /label=LC79 /note="ccdBspa_R" primer_bind complement(1833..1850) /label=LC99 /note="OriAspa_R" primer_bind 1904..1923 /label=B248 /note="VR" primer_bind 1911..1971 /label=LC75_CRRNAcos_R misc_feature 1951..2174 /label=Minimal lambda cos sequence primer_bind 1951..1971 /label=LC91 /note="Cas9cos_R" primer_bind 2074..2089 /label=B160 /note="rpsLsp-R" primer_bind complement(2084..2099) /label=LC118 /note="treF_F" misc_feature 2086..2097 /label=Lambda cos site primer_bind complement(2153..2214) /label=LC74_CRRNAcos_F primer_bind complement(2153..2174) /label=LC90 /note="Cas9cos_F" terminator 2189..2275 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 2272..2291 /label=LC20 /note="Ku-Seq7-R" primer_bind complement(2287..2305) /label=LC41 /note="ELZ2mutBB_F" primer_bind complement(2288..2304) /label=LC11 /note="pZS24-BB-F" primer_bind 2289..2313 /label=LC102 /note="pITCRRNA_F" primer_bind 2299..2329 /label=B298 - BamHI-leader-F /note="from pACYC184" CDS 2305..2334 /codon_start=2 /locus_tag="SPy_1049" /old_locus_tag="SPy1049" /product="hypothetical protein" /label='SPy_1049 /note="Best Blastp hit = pir||T03910 UDP-glucuronosyltransferase homolog F39G3.1 - Caenorhabditis elegans >gi|2291205|gb|AAB65327.1| (AF016424) Similar to UDP-glucosyltransferase [Caenorhabditis elegans]" /note="hypothetical protein" /db_xref="GI:13622196" /protein_id="AAK33939.1" /translation="IS**LKIWY" primer_bind 2358..2377 /label=D016 /note="pDB114 RSR F" primer_bind complement(2414..2436) /label=B220 /note="leader-sp1-R" primer_bind 2417..2467 /label=B690 /note="SpCRISPR-F" primer_bind 2426..2446 /label=B361 /note="(pTrc)-crispr-F" primer_bind complement(2428..2538) /label=D020 /note="Spa 2 R" primer_bind 2437..2532 /label=D003 /note="Spacer spa 1" misc_feature 2437..2468 /label=Repeat /note="Repeat" /note="Repeat" primer_bind 2437..2466 /label=D004 /note="Spacer spa 2" primer_bind 2437..2465 /label=D037 primer_bind 2437..2461 /label=B259 /note="bgaA-sp1-F" primer_bind 2437..2459 /label=D035 /note="phiNM4 F" primer_bind 2437..2459 /label=B217 /note="srtA-sp1-F" primer_bind 2437..2459 /label=B219 /note="sp1-rep-F" primer_bind 2437..2459 /label=B684 /note="(protAsp)rep-F" primer_bind 2437..2459 /label=D025 /note="GyrA F pDB269" primer_bind 2437..2459 /label=D029 /note="hla 1 F pDB269" primer_bind 2437..2459 /label=D031 /note="hla 2 F pDB269" primer_bind 2437..2459 /label=B223 /note="Pyo-BsaIsp-F" primer_bind 2437..2458 /label=B321 /note="bsaI-sp-F" primer_bind complement(2443..2472) /label=B691 /note="SpCRISPR-R" primer_bind complement(2444..2525) /label=D038 /note="GG hla 2 R" primer_bind complement(2448..2473) /label=B260 /note="bgaA-sp1-R" primer_bind complement(2449..2472) /label=B224 /note="Pyo-BsaIsp-R" primer_bind complement(2450..2473) /label=D032 /note="hla 2 R pDB269" primer_bind complement(2450..2473) /label=D026 /note="GyrA R pDB269" primer_bind complement(2450..2472) /label=D030 /note="hla 1 R pDB269" primer_bind complement(2450..2472) /label=B685 /note="(protAsp)rep-R" primer_bind complement(2450..2472) /label=B218 /note="srtA-sp1-R" primer_bind complement(2450..2472) /label=D036 /note="phiNM4 R" primer_bind 2451..2531 /label=D037 primer_bind complement(2451..2472) /label=D007 /note="spacer R" primer_bind complement(2451..2472) /label=B320 /note="bsaI-sp-R" primer_bind complement(2456..2474) /label=LC26 /note="ELZ1n2-R2" primer_bind complement(2456..2473) /label=B639 /note="mecA_dbsp3-R" primer_bind complement(2456..2472) /label=B657 /note="pUSA2_2sp" primer_bind complement(2456..2472) /label=B641 /note="sek_dbsp-R" primer_bind 2502..2533 /label=B690 /note="SpCRISPR-F" primer_bind 2503..2614 /label=D018 /note="Spa 1 F" primer_bind 2503..2614 /label=D019 /note="Spa 2 F" primer_bind 2503..2614 /label=D021 /note="arlR F" primer_bind 2503..2614 /label=D022 /note="arlS F" primer_bind 2503..2598 /label=D003 /note="Spacer spa 1" primer_bind 2503..2598 /label=D004 /note="Spacer spa 2" primer_bind 2503..2527 /label=B259 /note="bgaA-sp1-F" primer_bind 2503..2525 /label=D035 /note="phiNM4 F" primer_bind 2503..2525 /label=B217 /note="srtA-sp1-F" primer_bind 2503..2525 /label=B219 /note="sp1-rep-F" primer_bind 2503..2525 /label=B684 /note="(protAsp)rep-F" primer_bind 2503..2525 /label=D025 /note="GyrA F pDB269" primer_bind 2503..2525 /label=D029 /note="hla 1 F pDB269" primer_bind 2503..2525 /label=D031 /note="hla 2 F pDB269" primer_bind 2503..2525 /label=B223 /note="Pyo-BsaIsp-F" primer_bind 2503..2524 /label=B321 /note="bsaI-sp-F" primer_bind complement(2509..2538) /label=B691 /note="SpCRISPR-R" primer_bind complement(2510..2591) /label=D038 /note="GG hla 2 R" primer_bind complement(2514..2573) /label=B260 /note="bgaA-sp1-R" primer_bind complement(2515..2540) /label=B224 /note="Pyo-BsaIsp-R" primer_bind complement(2516..2541) /label=D030 /note="hla 1 R pDB269" primer_bind complement(2516..2539) /label=D036 /note="phiNM4 R" primer_bind complement(2516..2538) /label=D032 /note="hla 2 R pDB269" primer_bind complement(2516..2538) /label=D026 /note="GyrA R pDB269" primer_bind complement(2516..2538) /label=B685 /note="(protAsp)rep-R" primer_bind complement(2516..2538) /label=B218 /note="srtA-sp1-R" primer_bind 2517..2597 /label=D037 primer_bind complement(2517..2538) /label=D007 /note="spacer R" primer_bind complement(2517..2538) /label=B320 /note="bsaI-sp-R" primer_bind complement(2522..2540) /label=B657 /note="pUSA2_2sp" primer_bind complement(2522..2538) /label=LC26 /note="ELZ1n2-R2" primer_bind complement(2522..2538) /label=B641 /note="sek_dbsp-R" primer_bind complement(2522..2538) /label=B639 /note="mecA_dbsp3-R" primer_bind 2568..2591 /label=D031 /note="hla 2 F pDB269" primer_bind 2568..2591 /label=B223 /note="Pyo-BsaIsp-F" primer_bind 2569..2599 /label=B690 /note="SpCRISPR-F" primer_bind 2569..2593 /label=B259 /note="bgaA-sp1-F" primer_bind 2569..2591 /label=D035 /note="phiNM4 F" primer_bind 2569..2591 /label=B217 /note="srtA-sp1-F" primer_bind 2569..2591 /label=B219 /note="sp1-rep-F" primer_bind 2569..2591 /label=B684 /note="(protAsp)rep-F" primer_bind 2569..2591 /label=D025 /note="GyrA F pDB269" primer_bind 2569..2591 /label=D029 /note="hla 1 F pDB269" primer_bind 2569..2590 /label=B321 /note="bsaI-sp-F" misc_feature 2570..2604 /label=Repeat /note="Repeat" /note="Repeat" primer_bind complement(2575..2625) /label=B691 /note="SpCRISPR-R" primer_bind complement(2576..2604) /label=D038 /note="GG hla 2 R" primer_bind 2580..2620 /label=B374 /note="rep_mut-F" primer_bind complement(2580..2604) /label=B260 /note="bgaA-sp1-R" primer_bind complement(2581..2604) /label=B224 /note="Pyo-BsaIsp-R" primer_bind complement(2582..2605) /label=B218 /note="srtA-sp1-R" primer_bind complement(2582..2604) /label=D032 /note="hla 2 R pDB269" primer_bind complement(2582..2604) /label=D030 /note="hla 1 R pDB269" primer_bind complement(2582..2604) /label=D026 /note="GyrA R pDB269" primer_bind complement(2582..2604) /label=B685 /note="(protAsp)rep-R" primer_bind complement(2582..2604) /label=D036 /note="phiNM4 R" primer_bind complement(2583..2605) /label=B320 /note="bsaI-sp-R" primer_bind complement(2583..2604) /label=D007 /note="spacer R" primer_bind complement(2588..2604) /label=LC26 /note="ELZ1n2-R2" primer_bind complement(2588..2604) /label=B657 /note="pUSA2_2sp" primer_bind complement(2588..2604) /label=B641 /note="sek_dbsp-R" primer_bind complement(2588..2604) /label=B639 /note="mecA_dbsp3-R" primer_bind 2604..2626 /label=Tracr F primer_bind 2605..2667 /label=LC35_tracrRNA_R misc_feature 2605..2639 /label=pre-tracr sequence primer_bind complement(2607..2639) /label=B299 - EcoRI-rep-R /note="from pACYC184" primer_bind complement(2607..2630) /label=B527 /note="SphI-CRISPR-R" primer_bind complement(2607..2630) /label=B333 /note="(cat)-CRISPR-R" primer_bind complement(2607..2630) /label=B328 /note="(pC194)-cas9-R" misc_feature 2640..2810 /label=tracrRNA /note="tracrRNA" /note="tracrRNA" primer_bind 2723..2744 /label=B260 /note="bgaA-sp1-R" primer_bind 2723..2743 /label=B224 /note="Pyo-BsaIsp-R" primer_bind 2723..2742 /label=D032 /note="hla 2 R pDB269" primer_bind 2723..2742 /label=D030 /note="hla 1 R pDB269" primer_bind 2723..2742 /label=D026 /note="GyrA R pDB269" primer_bind 2723..2742 /label=B685 /note="(protAsp)rep-R" primer_bind 2723..2742 /label=B218 /note="srtA-sp1-R" primer_bind 2723..2742 /label=D036 /note="phiNM4 R" primer_bind 2723..2741 /label=D007 /note="spacer R" primer_bind 2723..2741 /label=B320 /note="bsaI-sp-R" primer_bind complement(2731..2747) /label=B259 /note="bgaA-sp1-F" primer_bind complement(2733..2747) /label=D035 /note="phiNM4 F" primer_bind complement(2733..2747) /label=B217 /note="srtA-sp1-F" primer_bind complement(2733..2747) /label=B219 /note="sp1-rep-F" primer_bind complement(2733..2747) /label=B684 /note="(protAsp)rep-F" primer_bind complement(2733..2747) /label=D025 /note="GyrA F pDB269" primer_bind complement(2733..2747) /label=D029 /note="hla 1 F pDB269" primer_bind complement(2733..2747) /label=D031 /note="hla 2 F pDB269" primer_bind complement(2733..2747) /label=B223 /note="Pyo-BsaIsp-F" primer_bind complement(2734..2747) /label=B321 /note="bsaI-sp-F" misc_feature 2811..2840 /label=post-tracr sequence primer_bind complement(2818..2842) /label=dCas9-tracr R primer_bind complement(2818..2840) /label=Tracr R primer_bind 2912..2934 /label=B254 /note="tracrRNA-R" primer_bind complement(2916..2935) /label=LC103 /note="pITCRRNA_R" primer_bind complement(2919..22) /label=LC36_tracrRNA_F primer_bind 2919..22 /label=LC34_pCRISPR_BB_R ORIGIN 1 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt 61 ttcttagacg tcggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa 121 agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga 181 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc 241 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg 301 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac 361 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc 421 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg 481 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga 541 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg 601 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg 661 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt 721 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc 781 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg 841 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga 901 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc 961 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc 1021 gagagctcgc ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat 1081 ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagcactagt 1141 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa 1201 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 1261 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc 1321 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta 1381 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 1441 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 1501 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 1561 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc 1621 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 1681 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 1741 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 1801 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca 1861 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg 1921 agctgatacc gctcgccgca gccgaacgcc ccaaaaagcc tcgctttcag cacctgtcgt 1981 ttcctttctt ttcagagggt attttaaata aaaacattaa gttatgacga agaagaacgg 2041 aaacgcctta aaccggaaaa ttttcataaa tagcgaaaac ccgcgaggtc gccgccccgt 2101 aacctgtcgg atcaccggaa aggacccgta aagtgataat gattatcatc tacatatcac 2161 aacgtgcgtg gagggtctag ggcggcggat ttgtcctact caggagagcg ttcaccgaca 2221 aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc 2281 tctagcacgc gtaccatggg atcctatttc ttaataacta aaaatatggt ataatactct 2341 taataaatgc agtaatacag gggcttttca agactgaagt ctagctgaga caaatagtgc 2401 gattacgaaa ttttttagac aaaaatagtc tacgaggttt tagagctatg ctgttttgaa 2461 tggtcccaaa acccttcacc ttgtggtgct tcagagaatt tggttttaga gctatgctgt 2521 tttgaatggt cccaaaacag tttggcggag acccacgctt catacggagt tttagagcta 2581 tgctgttttg aatggtccca aaacttcagc acactgagac ttgttgagtt gaattcatga 2641 gtattaagta ttgttttatg gctgataaat ttctttgaat ttctccttga ttatttgtta 2701 taaaagttat aaaataatct tgttggaacc attcaaaaca gcatagcaag ttaaaataag 2761 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt gatacttcta 2821 ttctactctg actgcaaacc aaaaaaacaa gcgctttcaa aacgcttgtt ttatcatttt 2881 tagggaaatt aatctcttaa tccttttatc attctacatt taggcgctgc catctt //