LOCUS       Exported                2936 bp ds-DNA     circular SYN 18-JUL-2016
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     2..21
                     /label=LC103
                     /note="pITCRRNA_R"
     primer_bind     complement(3..25)
                     /label=B254
                     /note="tracrRNA-R"
     primer_bind     95..119
                     /label=dCas9-tracr R
     misc_feature    complement(97..126)
                     /label=post-tracr sequence
     primer_bind     97..119
                     /label=Tracr R
     misc_feature    complement(127..297)
                     /label=tracrRNA
                     /note="tracrRNA"
                     /note="tracrRNA"
     primer_bind     190..206
                     /label=B259
                     /note="bgaA-sp1-F"
     primer_bind     190..204
                     /label=D035
                     /note="phiNM4 F"
     primer_bind     190..204
                     /label=B217
                     /note="srtA-sp1-F"
     primer_bind     190..204
                     /label=B219
                     /note="sp1-rep-F"
     primer_bind     190..204
                     /label=B684
                     /note="(protAsp)rep-F"
     primer_bind     190..204
                     /label=D025
                     /note="GyrA F pDB269"
     primer_bind     190..204
                     /label=D029
                     /note="hla 1 F pDB269"
     primer_bind     190..204
                     /label=D031
                     /note="hla 2 F pDB269"
     primer_bind     190..204
                     /label=B223
                     /note="Pyo-BsaIsp-F"
     primer_bind     190..203
                     /label=B321
                     /note="bsaI-sp-F"
     primer_bind     complement(193..214)
                     /label=B260
                     /note="bgaA-sp1-R"
     primer_bind     complement(194..214)
                     /label=B224
                     /note="Pyo-BsaIsp-R"
     primer_bind     complement(195..214)
                     /label=D032
                     /note="hla 2 R pDB269"
     primer_bind     complement(195..214)
                     /label=D030
                     /note="hla 1 R pDB269"
     primer_bind     complement(195..214)
                     /label=D026
                     /note="GyrA R pDB269"
     primer_bind     complement(195..214)
                     /label=B685
                     /note="(protAsp)rep-R"
     primer_bind     complement(195..214)
                     /label=B218
                     /note="srtA-sp1-R"
     primer_bind     complement(195..214)
                     /label=D036
                     /note="phiNM4 R"
     primer_bind     complement(196..214)
                     /label=D007
                     /note="spacer R"
     primer_bind     complement(196..214)
                     /label=B320
                     /note="bsaI-sp-R"
     primer_bind     complement(270..332)
                     /label=LC35_tracrRNA_R
     misc_feature    complement(298..332)
                     /label=pre-tracr sequence
     primer_bind     298..330
                     /label=B299 - EcoRI-rep-R
                     /note="from pACYC184"
     primer_bind     307..330
                     /label=B527
                     /note="SphI-CRISPR-R"
     primer_bind     307..330
                     /label=B333
                     /note="(cat)-CRISPR-R"
     primer_bind     307..330
                     /label=B328
                     /note="(pC194)-cas9-R"
     primer_bind     complement(311..333)
                     /label=Tracr F
     primer_bind     312..362
                     /label=B691
                     /note="SpCRISPR-R"
     primer_bind     complement(317..357)
                     /label=B374
                     /note="rep_mut-F"
     primer_bind     complement(323..434)
                     /label=D018
                     /note="Spa 1 F"
     primer_bind     complement(323..434)
                     /label=D019
                     /note="Spa 2 F"
     primer_bind     complement(323..434)
                     /label=D021
                     /note="arlR F"
     primer_bind     complement(323..434)
                     /label=D022
                     /note="arlS F"
     primer_bind     332..355
                     /label=B218
                     /note="srtA-sp1-R"
     primer_bind     332..354
                     /label=B320
                     /note="bsaI-sp-R"
     misc_feature    complement(333..363)
                     /label=Repeat
                     /note="Repeat"
                     /note="Repeat"
     primer_bind     333..361
                     /label=D038
                     /note="GG hla 2 R"
     primer_bind     333..357
                     /label=B260
                     /note="bgaA-sp1-R"
     primer_bind     333..356
                     /label=B224
                     /note="Pyo-BsaIsp-R"
     primer_bind     333..355
                     /label=D032
                     /note="hla 2 R pDB269"
     primer_bind     333..355
                     /label=D030
                     /note="hla 1 R pDB269"
     primer_bind     333..355
                     /label=D026
                     /note="GyrA R pDB269"
     primer_bind     333..355
                     /label=B685
                     /note="(protAsp)rep-R"
     primer_bind     333..355
                     /label=D036
                     /note="phiNM4 R"
     primer_bind     333..354
                     /label=D007
                     /note="spacer R"
     primer_bind     333..349
                     /label=LC26
                     /note="ELZ1n2-R2"
     primer_bind     333..349
                     /label=B657
                     /note="pUSA2_2sp"
     primer_bind     333..349
                     /label=B641
                     /note="sek_dbsp-R"
     primer_bind     333..349
                     /label=B639
                     /note="mecA_dbsp3-R"
     primer_bind     complement(338..368)
                     /label=B690
                     /note="SpCRISPR-F"
     primer_bind     complement(339..434)
                     /label=D003
                     /note="Spacer spa 1"
     primer_bind     complement(339..434)
                     /label=D004
                     /note="Spacer spa 2"
     primer_bind     complement(340..420)
                     /label=D037
     primer_bind     complement(344..368)
                     /label=B259
                     /note="bgaA-sp1-F"
     primer_bind     346..427
                     /label=D038
                     /note="GG hla 2 R"
     primer_bind     complement(346..369)
                     /label=D031
                     /note="hla 2 F pDB269"
     primer_bind     complement(346..369)
                     /label=B223
                     /note="Pyo-BsaIsp-F"
     primer_bind     complement(346..368)
                     /label=D035
                     /note="phiNM4 F"
     primer_bind     complement(346..368)
                     /label=B217
                     /note="srtA-sp1-F"
     primer_bind     complement(346..368)
                     /label=B219
                     /note="sp1-rep-F"
     primer_bind     complement(346..368)
                     /label=B684
                     /note="(protAsp)rep-F"
     primer_bind     complement(346..368)
                     /label=D025
                     /note="GyrA F pDB269"
     primer_bind     complement(346..368)
                     /label=D029
                     /note="hla 1 F pDB269"
     primer_bind     complement(347..368)
                     /label=B321
                     /note="bsaI-sp-F"
     primer_bind     396..421
                     /label=D030
                     /note="hla 1 R pDB269"
     primer_bind     397..422
                     /label=B224
                     /note="Pyo-BsaIsp-R"
     primer_bind     397..415
                     /label=B657
                     /note="pUSA2_2sp"
     primer_bind     398..421
                     /label=D036
                     /note="phiNM4 R"
     primer_bind     399..509
                     /label=D020
                     /note="Spa 2 R"
     primer_bind     399..428
                     /label=B691
                     /note="SpCRISPR-R"
     primer_bind     399..423
                     /label=B260
                     /note="bgaA-sp1-R"
     primer_bind     399..421
                     /label=D032
                     /note="hla 2 R pDB269"
     primer_bind     399..421
                     /label=D026
                     /note="GyrA R pDB269"
     primer_bind     399..421
                     /label=B685
                     /note="(protAsp)rep-R"
     primer_bind     399..421
                     /label=B218
                     /note="srtA-sp1-R"
     primer_bind     399..420
                     /label=D007
                     /note="spacer R"
     primer_bind     399..420
                     /label=B320
                     /note="bsaI-sp-R"
     primer_bind     399..415
                     /label=LC26
                     /note="ELZ1n2-R2"
     primer_bind     399..415
                     /label=B641
                     /note="sek_dbsp-R"
     primer_bind     399..415
                     /label=B639
                     /note="mecA_dbsp3-R"
     primer_bind     complement(404..434)
                     /label=B690
                     /note="SpCRISPR-F"
     primer_bind     complement(405..500)
                     /label=D003
                     /note="Spacer spa 1"
     primer_bind     complement(406..486)
                     /label=D037
     primer_bind     complement(410..434)
                     /label=B259
                     /note="bgaA-sp1-F"
     primer_bind     412..493
                     /label=D038
                     /note="GG hla 2 R"
     primer_bind     complement(412..435)
                     /label=D035
                     /note="phiNM4 F"
     primer_bind     complement(412..435)
                     /label=B219
                     /note="sp1-rep-F"
     primer_bind     complement(412..435)
                     /label=B684
                     /note="(protAsp)rep-F"
     primer_bind     complement(412..435)
                     /label=D029
                     /note="hla 1 F pDB269"
     primer_bind     complement(412..434)
                     /label=B217
                     /note="srtA-sp1-F"
     primer_bind     complement(412..434)
                     /label=D025
                     /note="GyrA F pDB269"
     primer_bind     complement(412..434)
                     /label=D031
                     /note="hla 2 F pDB269"
     primer_bind     complement(412..434)
                     /label=B223
                     /note="Pyo-BsaIsp-F"
     primer_bind     complement(413..434)
                     /label=B321
                     /note="bsaI-sp-F"
     primer_bind     complement(421..434)
                     /label=LC23
                     /note="ELZ1n2-F1"
     primer_bind     462..494
                     /label=B691
                     /note="SpCRISPR-R"
     primer_bind     464..488
                     /label=B224
                     /note="Pyo-BsaIsp-R"
     primer_bind     464..487
                     /label=D030
                     /note="hla 1 R pDB269"
     primer_bind     464..487
                     /label=D036
                     /note="phiNM4 R"
     primer_bind     464..481
                     /label=B657
                     /note="pUSA2_2sp"
     misc_feature    complement(465..500)
                     /label=Repeat
                     /note="Repeat"
                     /note="Repeat"
     primer_bind     465..489
                     /label=B260
                     /note="bgaA-sp1-R"
     primer_bind     465..487
                     /label=D032
                     /note="hla 2 R pDB269"
     primer_bind     465..487
                     /label=D026
                     /note="GyrA R pDB269"
     primer_bind     465..487
                     /label=B685
                     /note="(protAsp)rep-R"
     primer_bind     465..487
                     /label=B218
                     /note="srtA-sp1-R"
     primer_bind     465..486
                     /label=D007
                     /note="spacer R"
     primer_bind     465..486
                     /label=B320
                     /note="bsaI-sp-R"
     primer_bind     465..481
                     /label=LC26
                     /note="ELZ1n2-R2"
     primer_bind     465..481
                     /label=B641
                     /note="sek_dbsp-R"
     primer_bind     465..481
                     /label=B639
                     /note="mecA_dbsp3-R"
     primer_bind     complement(470..520)
                     /label=B690
                     /note="SpCRISPR-F"
     primer_bind     complement(471..500)
                     /label=D004
                     /note="Spacer spa 2"
     primer_bind     complement(472..500)
                     /label=D037
     primer_bind     complement(476..500)
                     /label=B259
                     /note="bgaA-sp1-F"
     primer_bind     complement(478..500)
                     /label=D035
                     /note="phiNM4 F"
     primer_bind     complement(478..500)
                     /label=B217
                     /note="srtA-sp1-F"
     primer_bind     complement(478..500)
                     /label=B219
                     /note="sp1-rep-F"
     primer_bind     complement(478..500)
                     /label=B684
                     /note="(protAsp)rep-F"
     primer_bind     complement(478..500)
                     /label=D025
                     /note="GyrA F pDB269"
     primer_bind     complement(478..500)
                     /label=D029
                     /note="hla 1 F pDB269"
     primer_bind     complement(478..500)
                     /label=D031
                     /note="hla 2 F pDB269"
     primer_bind     complement(478..500)
                     /label=B223
                     /note="Pyo-BsaIsp-F"
     primer_bind     complement(479..500)
                     /label=B321
                     /note="bsaI-sp-F"
     primer_bind     complement(491..511)
                     /label=B361
                     /note="(pTrc)-crispr-F"
     primer_bind     501..523
                     /label=B220
                     /note="leader-sp1-R"
     primer_bind     complement(560..579)
                     /label=D016
                     /note="pDB114 RSR F"
     CDS             complement(603..632)
                     /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     complement(608..638)
                     /label=B298 - BamHI-leader-F
                     /note="from pACYC184"
     primer_bind     complement(624..648)
                     /label=LC102
                     /note="pITCRRNA_F"
     primer_bind     632..650
                     /label=LC41
                     /note="ELZ2mutBB_F"
     primer_bind     633..649
                     /label=LC11
                     /note="pZS24-BB-F"
     primer_bind     complement(646..665)
                     /label=LC20
                     /note="Ku-Seq7-R"
     terminator      662..748
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     723..784
                     /label=LC74_CRRNAcos_F
     misc_feature    763..986
                     /label=Minimal lambda cos sequence
     primer_bind     763..784
                     /label=LC90
                     /note="Cas9cos_F"
     primer_bind     838..853
                     /label=LC118
                     /note="treF_F"
     misc_feature    840..851
                     /label=Lambda cos site
     primer_bind     complement(848..863)
                     /label=B160
                     /note="rpsLsp-R"
     primer_bind     complement(966..1026)
                     /label=LC75_CRRNAcos_R
     primer_bind     complement(966..986)
                     /label=LC91
                     /note="Cas9cos_R"
     primer_bind     complement(1014..1033)
                     /label=B248
                     /note="VR"
     primer_bind     1087..1104
                     /label=LC99
                     /note="OriAspa_R"
     rep_origin      complement(1133..1721)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1633..1652
                     /label=LC79
                     /note="ccdBspa_R"
     terminator      1809..1903
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    1906..1935
                     /label=CBB CRISPR spapcer
                     /note="cut pELZ2mut1K backbone, cut pCRISPR, pZE21, pCRRNA 
                     backbone
                     "
     CDS             complement(1934..2728)
                     /codon_start=1
                     /label=kanR
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     2812..2834
                     /label=B385
     primer_bind     2821..2843
                     /label=B377 pZ-F
                     /note="from pDB129 pZE21-CRISPR-bsaI"
     primer_bind     complement(2845..2864)
                     /label=LC13
                     /note="pZS24-BB-R"
     primer_bind     2915..18
                     /label=LC36_tracrRNA_F
     primer_bind     complement(2915..18)
                     /label=LC34_pCRISPR_BB_R
     primer_bind     complement(2919..2936)
                     /label=LC42
                     /note="ELZ2MutBB_R"
ORIGIN
        1 aagatggcag cgcctaaatg tagaatgata aaaggattaa gagattaatt tccctaaaaa
       61 tgataaaaca agcgttttga aagcgcttgt ttttttggtt tgcagtcaga gtagaataga
      121 agtatcaaaa aaagcaccga ctcggtgcca ctttttcaag ttgataacgg actagcctta
      181 ttttaacttg ctatgctgtt ttgaatggtt ccaacaagat tattttataa cttttataac
      241 aaataatcaa ggagaaattc aaagaaattt atcagccata aaacaatact taatactcat
      301 gaattcaact caacaagtct cagtgtgctg aagttttggg accattcaaa acagcatagc
      361 tctaaaactc cgtatgaagg cacccactcc gccaaactgt tttgggacca ttcaaaacag
      421 catagctcta aaacactctt aatgtcgcga ccaggatgtg cgatgttttg ggaccattca
      481 aaacagcata gctctaaaac ctcgtagact atttttgtct aaaaaatttc gtaatcgcac
      541 tatttgtctc agctagactt cagtcttgaa aagcccctgt attactgcat ttattaagag
      601 tattatacca tatttttagt tattaagaaa taggatccca tggtacgcgt gctagaggca
      661 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
      721 ggtgaacgct ctcctgagta ggacaaatcc gccgccctag accctccacg cacgttgtga
      781 tatgtagatg ataatcatta tcactttacg ggtcctttcc ggtgatccga caggttacgg
      841 ggcggcgacc tcgcgggttt tcgctattta tgaaaatttt ccggtttaag gcgtttccgt
      901 tcttcttcgt cataacttaa tgtttttatt taaaataccc tctgaaaaga aaggaaacga
      961 caggtgctga aagcgaggct ttttggggcg ttcggctgcg gcgagcggta tcagctcact
     1021 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1081 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1141 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1201 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1261 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1321 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1381 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1441 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1501 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1561 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1621 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1681 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1741 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgacta
     1801 gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
     1861 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga gctctcgaac
     1921 cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
     1981 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
     2041 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
     2101 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     2161 cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca
     2221 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     2281 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     2341 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     2401 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     2461 cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca
     2521 gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct
     2581 tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc
     2641 cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac
     2701 ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag
     2761 atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc
     2821 agggcttccc aaccttacca gagggcgccc cagctggcaa ttccgacgtc taagaaacca
     2881 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctt
//
