LOCUS       Exported               11833 bp ds-DNA     circular SYN 06-JUL-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11833)
  AUTHORS   Nishida
  TITLE     Direct Submission
  JOURNAL   Exported Jul 6, 2022 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11833
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..36
                     /label=kn502
                     /note="pRSend_Sph1_R"
                     /note="color: ?; sequence: 
                     gacgaaagggcctcggcatgcgcctatttttatagg"
     primer_bind     complement(1..36)
                     /label=kn501
                     /note="pRSend_Sph1_F"
                     /note="color: ?; sequence: 
                     cctataaaaataggcgcatgccgaggccctttcgtc"
     primer_bind     8..23
                     /label=kn1219
                     /note="Ura-Ade-Sma-19_R"
                     /note="color: ?; sequence: 
                     CAATGCTTATGATTCGATTGTTCTTAATAGTACCAACTGCAAAAGATAAAACCCGGGCC
                     TCGTGATACGC"
     primer_bind     54..74
                     /label=kn2480
                     /note="Sph-Nsi-CEN_F"
                     /note="color: ?; sequence: 
                     cgtgcatgcatgcatggtttcttaggacggatcg"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
                     /note="color: #454f59"
     primer_bind     70..90
                     /label=kn1589
                     /note="Km_gRNAvector_1"
                     /note="color: ?; sequence: 
                     CGTtctagatggtacctgctagctggtaaccgatcgcttgcctgtaacttac"
     primer_bind     582..604
                     /label=kn2519
                     /note="Amp-XmaI_F"
                     /note="color: ?; sequence: 
                     gaagtcagccccgggtggcacttttcggggaa"
     primer_bind     582..600
                     /label=kn909
                     /note="aatL2-amp_F"
                     /note="color: ?; sequence: 
                     ttcttgtacaaagttgggcggccgcggatcccaggtggcacttttcgg"
     primer_bind     583..601
                     /label=hm020
                     /note="pET24 downKm ga FOR,42"
                     /note="color: ?; sequence: CCAGAGCTGCcaggtggcacttttcggg"
     primer_bind     584..602
                     /label=hm387
                     /note="amp pro for ga btm,36"
                     /note="color: ?; sequence: 
                     ATCTGGCCCGTACATCGCAGGTGGCACTTTTCGGGG"
     primer_bind     584..602
                     /label=hm029
                     /note="rrn-downKm ga FOR,38"
                     /note="color: ?; sequence: 
                     GACGGATGGCCTTTTTGCGcaggtggcacttttcgggg"
     primer_bind     584..602
                     /label=hm545
                     /note="Amp-ori casette for ga w T7-MS2 top,39"
                     /note="color: ?; sequence: 
                     gggtcttgaggggttttttgCAGGTGGCACTTTTCGGGG"
     primer_bind     584..598
                     /label=kn853
                     /note="rrnBt-Nsi-Aat2_R"
                     /note="color: ?; sequence: 
                     GGCCTTTCGTTTTATctgttgtttgCGGCCGatcatgcatgagacgtcaggtggcactt
                     ttc"
     primer_bind     586..603
                     /label=kn2073
                     /note="AgeI-H1p_R"
                     /note="color: ?; sequence: 
                     GACGTCAGCGTTCGAATTggtacaccggtggcacttttcgggGA"
     primer_bind     complement(589..613)
                     /label=kn1154
                     /note="pKNT19_aat2_F"
                     /note="color: ?; sequence: 
                     cctGTGTATAACTCACTACTTTAGTCAGTTCCgacgtgcACATTTCCCCGAAAAGTGCC
                     "
     primer_bind     complement(591..616)
                     /label=hm780
                     /note="gRNA cassette btm for GA w/ pET_Ago+MF,39"
                     /note="color: ?; sequence: 
                     gcatcaggcgctcttccgctCACATTTCCCCGAAAAGTG"
     primer_bind     609..626
                     /label=hm665
                     /note="AmpR pro upstream top,38"
                     /note="color: ?; sequence: 
                     ctgtcgcacaaccatgGCTAGcgcggaacccctatttg"
     primer_bind     609..626
                     /label=kn2434
                     /note="Leu-IsceAvr-Amp_F"
                     /note="color: ?; sequence: 
                     cacattaccctgttatccctaggcgcggaacccctatttg"
     promoter        610..714
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     610..626
                     /label=hm517
                     /note="AmpR pro 5' fw for seq,17"
                     /note="color: ?; sequence: cgcggaacccctatttg"
     primer_bind     627..647
                     /label=hm723
                     /note="KmR dw btm,36"
                     /note="color: ?; sequence: 
                     ctaaaagccagataaTTTATTTTTCTAAATACATTC"
     primer_bind     complement(658..677)
                     /label=kn708
                     /note="AmpP_R"
                     /note="color: ?; sequence: ggttattgtctcatgagcgg"
     CDS             715..1575
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1:  715 ..  783 / #ccffcc / signal sequence
                        2:  784 .. 1575 / #ccffcc
                     Cleavage site after base 783"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     715..734
                     /label=kn397
                     /note="B-lactamase_F"
                     /note="color: ?; sequence: ATGAGTATTCAACATTTCCG"
     primer_bind     complement(757..776)
                     /label=kn398
                     /note="B-Lactamase_50_R"
                     /note="color: ?; sequence: ACAGGAAGGCAAAATGCCGC"
     primer_bind     complement(782..803)
                     /label=hm518
                     /note="AmpR 30th aa rv for seq,22"
                     /note="color: ?; sequence: ttcaccagcgtttctgggtgag"
     primer_bind     789..811
                     /label=kn184
                     /note="BLF1_ENX_F"
                     /note="color: ?; sequence: 
                     tgaattcgcggccgcttctagaGAAACGCTGGTGAAAGTAAAAG"
     primer_bind     complement(1136..1155)
                     /label=hm656
                     /note="AmpR mdl btm,20"
                     /note="color: ?; sequence: ggttagctccttcggtcctc"
     primer_bind     complement(1275..1296)
                     /label=kn185
                     /note="BLF1_SNP_R"
                     /note="color: ?; sequence: 
                     ctgcagcggccgctactagtGCCAGTTAATAGTTTGCGCaac"
     primer_bind     1299..1319
                     /label=kn188
                     /note="BLF2_ENX_F"
                     /note="color: ?; sequence: 
                     tgaattcgcggccgcttctagaCTACTTACTCTAGCTTCCCG"
     primer_bind     1350..1373
                     /label=hm519
                     /note="AmpR 212nd aa fw for seq,24"
                     /note="color: ?; sequence: taaagttgcaggaccacttctgcg"
     primer_bind     complement(1416..1447)
                     /label=kn704
                     /note="ampR-Bsa1KO_R"
                     /note="color: ?; sequence: 
                     caatgataccgcgagaaccacgctcaccggct"
     primer_bind     1421..1450
                     /label=kn705
                     /note="ampR-Bsa1KO_F"
                     /note="color: ?; sequence: gtgagcgtggttctcgcggtatcattgcag"
     primer_bind     1507..1526
                     /label=hm516
                     /note="AmpR 265th aa fw for seq,20"
                     /note="color: ?; sequence: caggcaactatggatgaacg"
     primer_bind     complement(1553..1572)
                     /label=kn189
                     /note="BLF2_SNP_R"
                     /note="color: ?; sequence: 
                     ctgcagcggccgctactagtCCAATGCTTAATCAGTGAGG"
     primer_bind     complement(1653..1677)
                     /label=kn976
                     /note="amp-age_R"
                     /note="color: ?; sequence: 
                     catgatcactagatcctaggactctagattaccggtcatgagattatcaaaaaggatc"
     primer_bind     complement(1656..1681)
                     /label=kn874
                     /note="AmpAgeXbaChimr_F"
                     /note="color: ?; sequence: 
                     CCtctagactaccggttttggtcatgagattatcaaaaagg"
     primer_bind     complement(1657..1683)
                     /label=kn2520
                     /note="Amp-Cla_R"
                     /note="color: ?; sequence: 
                     cgcaacgcaatCgattttggtcatgagattatcaaaaag"
     primer_bind     complement(1657..1674)
                     /label=hm752
                     /note="Amp-ori for ga with 1105 fw,41"
                     /note="color: ?; sequence: 
                     TATGGCGTGCTGCTAGCcgatggcatgagattatcaaaaag"
     primer_bind     complement(1663..1691)
                     /label=kn879
                     /note="ampT-age1-p15A_R"
                     /note="color: ?; sequence: 
                     CATCTTCCAGGAAATCTCcgttaagggataccggtcatgagattatc"
     primer_bind     1670..1691
                     /label=hm788
                     /note="p15A ori up,44"
                     /note="color: ?; sequence: 
                     TGTAAGCAGACAGTTTTATTGTTCATGACCAAAATCCCTTAACG"
     primer_bind     complement(1677..1699)
                     /label=kn2547
                     /note="BsaX-DR_F"
                     /note="color: ?; sequence: 
                     gCcggagnacgcgtgcGTTTAAGAGCTATGCTGTTTTGAATGGTCCCAAAACTCACGTT
                     AAGGGATTTTG"
     primer_bind     1699..1726
                     /label=hm462
                     /note="cfSGFP-N vec for GA top,30"
                     /note="color: ?; sequence: gatcgttccactgagcgtcagaccccgtag"
     primer_bind     1707..1726
                     /label=hm892
                     /note="Cas9 gRNA Negative ctrl btm,25"
                     /note="color: ?; sequence: AAAACCTGAGCGTCAGACCCCGTAG"
     primer_bind     1707..1722
                     /label=kn2546
                     /note="23100up_R"
                     /note="color: ?; sequence: 
                     TAGCAcTgTACCTAGGACTGAGCTAGCCGTCAAGAGGAAATTTAAAATAATTTTCTGAG
                     CGTCAGACCCC"
     primer_bind     1725..1748
                     /label=kn875
                     /note="ChimerSpeNotColE_R"
                     /note="color: ?; sequence: 
                     GGTCATTAAAACtagtagcgccggcgagaaaagatcaaaggatcttcttg"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     1746..1767
                     /label=kn2443
                     /note="ori_R"
                     /note="color: ?; sequence: ttgagatcctttttttctgcgc"
     primer_bind     complement(1802..1821)
                     /label=kn2444
                     /note="ori_F"
                     /note="color: ?; sequence: caaacaaaccaccgctggta"
     primer_bind     complement(1873..1911)
                     /label=hm732
                     /note="ColE1 ori for C>T mut btm,39"
                     /note="color: ?; sequence: 
                     CTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGC"
     primer_bind     1894..1911
                     /label=hm733
                     /note="ColE1 ori for C>T mut top,18"
                     /note="color: ?; sequence: CTTCTAGTGTAGCCGTAG"
     primer_bind     2275..2293
                     /label=kn709
                     /note="ColE1"
                     /note="color: ?; sequence: cacctctgacttgagcgtc"
     primer_bind     complement(2312..2334)
                     /label=hm596
                     /note="ori 3' btm,23"
                     /note="color: ?; sequence: tttccataggctccgcccccctg"
     primer_bind     2318..2337
                     /label=kn746
                     /note="P15AendR"
                     /note="color: ?; sequence: GGCGGAGCCTATGGAAAAAC"
     primer_bind     complement(2318..2334)
                     /label=kn2433
                     /note="Ape-NheAat-ori_F"
                     /note="color: ?; sequence: 
                     ctCGCCTTAagtgctagcgacgtctaatttccataggctcggcc"
     primer_bind     complement(2319..2338)
                     /label=kn2403
                     /note="SNR-Spe-Ori_R"
                     /note="color: ?; sequence: 
                     caaagactagtggcgcgccgtttttccataggctccgc"
     primer_bind     complement(2319..2337)
                     /label=kn1590
                     /note="Km_gRNAvector_2"
                     /note="color: ?; sequence: 
                     aggccggccatagggataacagggtaatgttaaccggtttttccataggctccgc"
     primer_bind     complement(2337..2355)
                     /label=hm386
                     /note="sv40pa ori top for ga,36"
                     /note="color: ?; sequence: 
                     CGTCGACCTCTAGCTAGAAAGGCCGCGTTGCTGGCg"
     primer_bind     complement(2340..2358)
                     /label=kn2479
                     /note="ori-pacI_R"
                     /note="color: ?; sequence: 
                     ctgagtattcccacagttaattaaaaaggccgcgttgctg"
     primer_bind     complement(2350..2372)
                     /label=kn420
                     /note="Gal1R-pBR322_R"
                     /note="color: ?; sequence: 
                     GTATATGGATATGTATATGGTGGTAATGCCATGTAAGGCCAGGAACCGTAAAAAGGC"
     primer_bind     complement(2364..2383)
                     /label=hm593
                     /note="ori for HA-R,40"
                     /note="color: ?; sequence: 
                     ggggttcctggctctgctctAAGGCCAGCAAAAGGCCAGG"
     primer_bind     complement(2364..2383)
                     /label=hm620
                     /note="aavs1 HA I-SceI 1,58"
                     /note="color: ?; sequence: 
                     ggggttcctggctctgctctTAGGGATAACAGGGTAATAAGGCCAGCAAAAGGCCAGG"
     primer_bind     complement(2370..2388)
                     /label=kn1269
                     /note="pTDH3-Nhe-p426_R"
                     /note="color: ?; sequence: 
                     GGCAGTATTGATAATGAattcgctagcaaaaggccagcaaaag"
     primer_bind     complement(2375..2393)
                     /label=hm546
                     /note="Amp-ori casette for ga w T7-MS2 btm,40"
                     /note="color: ?; sequence: 
                     CTGGGCTGCTCGCCACAGGTTATGTGAGCAAAAGGCCAGC"
     primer_bind     complement(2380..2398)
                     /label=hm408
                     /note="upstream p15A ori in p909 top for ga,39"
                     /note="color: ?; sequence: 
                     GTTGTAAAACGACGGCCAGTAGAACATGTGAGCAAAAGG"
     primer_bind     complement(2380..2397)
                     /label=hm412
                     /note="Amp vec for hsvTK series GA btm,36"
                     /note="color: ?; sequence: 
                     GAAAGAACCAGCTGGGGCGAACATGTGAGCAAAAGG"
     primer_bind     2388..2406
                     /label=hm626
                     /note="CMV up top w dw bGHpA (hm463),39"
                     /note="color: ?; sequence: 
                     atcgttccactgagcgtcagtcacatgttctttcctgcg"
     primer_bind     complement(2413..2432)
                     /label=hm734
                     /note="ori down stream,20"
                     /note="color: ?; sequence: CGGTTATCCACAGAATCAGG"
     primer_bind     complement(2451..2468)
                     /label=kn1622
                     /note="pKNT19-ClaSpeNgo_F"
                     /note="color: ?; sequence: 
                     cTGCATCGATGAAGATTCTTGCTCactagtgcCGGCGAGCGGTATCAGCT"
     primer_bind     complement(2488..2507)
                     /label=kn707
                     /note="ColE-Bam-Spe-Sac1_R"
                     /note="color: ?; sequence: 
                     ATTTAAAAGCAGCGGGAGCTCagactagtcggatcctcgctcactgactcgct"
     primer_bind     complement(2495..2513)
                     /label=hm355
                     /note="sy5 Nco-frag2 to remove SapI top,29"
                     /note="color: ?; sequence: GGCGCTCgagTCCGCTTCCTCGCTCACTG"
     primer_bind     2515..2541
                     /label=hm354
                     /note="sy5 Nco-frag1 to remove SapI btm,35"
                     /note="color: ?; sequence: 
                     GCGGActcGAGCGCCCAATACGCAAACcgcctctc"
     primer_bind     complement(2560..2582)
                     /label=hm036
                     /note="pEX-A2J1 BsaI F,41"
                     /note="color: ?; sequence: 
                     tgaTCTAGAgagaccgggCTGTCGTGCCAGCTGCATTAATG"
     primer_bind     2596..2612
                     /label=kn2353
                     /note="LacI_end_F"
                     /note="color: ?; sequence: aaagcgggcagtgagcg"
     primer_bind     complement(2605..2621)
                     /label=kn2239
                     /note="LacI_OL_R"
                     /note="color: ?; sequence: 
                     gagataatttatcaccgcAGATGGTTATCGattgcgttgcgctcact"
     primer_bind     complement(2613..2633)
                     /label=kn1218
                     /note="ade1-HpaI-pUC19"
                     /note="color: ?; sequence: 
                     GTGGCCAGAGGTAAAGTTAGAGACATATATGAGGTAGACGCTGGTACGTTAACTCACAT
                     TAATTGCGTTG"
     primer_bind     2641..2662
                     /label=kn570
                     /note="T7t-Nsi1-LacP_F"
                     /note="color: ?; sequence: 
                     ttagaggccccaaggggttatgctatgcattaggcaccccaggctttac"
     primer_bind     2657..2693
                     /label=kn852
                     /note="LacO-Cas9_F"
                     /note="color: ?; sequence: 
                     ACTCATTATGGATTTAATTTAAAgggccctttacactttatgcttccggctcctatgtt
                     gtgtgg"
     promoter        2658..2688
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 4 segments:
                        1: 2658 .. 2663 / #ffffff / -35
                        2: 2664 .. 2680 / #ffffff
                        3: 2681 .. 2681 / #ffffff
                        4: 2682 .. 2688 / #ffffff / -10"
     primer_bind     complement(2670..2684)
                     /label=kn2488
                     /note="tracr-in-UV5_R"
                     /note="color: ?; sequence: 
                     AACTTGCTATGCTGTTTTGAATGGTTccAACAAGATCATTATACGAGCCGGAAGC"
     primer_bind     complement(2673..2712)
                     /label=kn851
                     /note="LacO-Cas9_R"
                     /note="color: ?; sequence: 
                     CCATTTTTGCCTCCTAAAATgggccttgttatccgctcacaattccacacaacatagga
                     gccgga"
     protein_bind    2696..2712
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #31849b"
     primer_bind     complement(2698..2714)
                     /label=hm070
                     /note="hm014 target 44-2,44"
                     /note="color: ?; sequence: 
                     CGAGGTAGTAGGTTGTAACTAGAGGGGAATTGTTATCCGCTCAC"
     primer_bind     2716..2737
                     /label=kn584
                     /note="M13R-Long"
                     /note="color: ?; sequence: cacacaggaaacagctatgacc"
     primer_bind     2719..2736
                     /label=kn98
                     /note="Pex3intR_M13R"
                     /note="color: ?; sequence: 
                     ggtaatgtcatgttacctattgcacacttactgtattaaagattacgctaCAGGAAACA
                     GCTATGAC"
     primer_bind     2719..2736
                     /label=kn99
                     /note="Pex19intR_M13R"
                     /note="color: ?; sequence: 
                     ccacttttttttttcattttttaaagtgttatacttagttatgctctaggataatgaac
                     taCAGGAAACAGCTATGAC"
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     2720..2736
                     /label=hm708
                     /note="M13 rev,17"
                     /note="color: ?; sequence: caggaaacagctatgac"
     primer_bind     complement(2731..2753)
                     /label=kn381
                     /note="LacZA1_R"
                     /note="color: ?; sequence: 
                     ACGACGGCCAGTGAGTTCGAGCTCGGAACCTCGCGAATACATCTGGATATCGGATCACG
                     GGCCCGTCGACTACAGAGGCCTGCATGCAAGCTTGGCGTAATCATGGTCATA"
     primer_bind     2731..2752
                     /label=kn413
                     /note="Left-LacZ_F"
                     /note="color: ?; sequence: 
                     ATACCACTGGCGGTGATACGTATGCATATGACCATGATTACGCCAAGC"
     primer_bind     2731..2746
                     /label=kn372
                     /note="cro-SD-LacZ_F"
                     /note="color: ?; sequence: 
                     GTAATAGAGGAGGTTAGTATGACCATGATTACG"
     primer_bind     complement(2733..2752)
                     /label=kn683
                     /note="NotBspEBcl-T3_R"
                     /note="color: ?; sequence: 
                     cgtccggattgcggccgcttggcgtaatcatggtca"
     primer_bind     2754..2779
                     /label=kn85
                     /note="JW412nest_R"
                     /note="color: ?; sequence: cgcaattaaccctcactaaagggaac"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     2766..2788
                     /label=kn682
                     /note="NotBspEBcl-T3_F"
                     /note="color: ?; sequence: 
                     ccgcaatccggacgtgatcactaaagggaacaaaagctgg"
     primer_bind     2768..2824
                     /label=kn2391
                     /note="Not-SHL_R"
                     /note="color: ?; sequence: 
                     actaaagggaacaaaagctgaccgggccatgcggccgcgatctccttctcttaggtggc
                     a"
     primer_bind     complement(2773..2817)
                     /label=kn1518
                     /note="SH3-LVA_F"
                     /note="color: ?; sequence: 
                     taagagaaggagagaattcgggcccggtcttgtggcataggtaccagcttttgttccct
                     "
     primer_bind     2788..2807
                     /label=kn770
                     /note="Kpn1-SHL_R"
                     /note="color: ?; sequence: gtaccgggcccgaattctct"
     primer_bind     complement(2794..2851)
                     /label=kn536_SH3_F
                     /note="color: ?; sequence: 
                     ctagtatggtcgaccctccacctgctctgccacctaagagaaggagagaattcgggcc"
     primer_bind     2798..2847
                     /label=kn537_SH3_R
                     /note="color: ?; sequence: 
                     cgaattctctccttctcttaggtggcagagcaggtggagggtcgaccata"
     misc_feature    2805..2837
                     /label=SH3 ligand
                     /note="color: #00ffff"
     primer_bind     2805..2825
                     /label=kn2359
                     /note="SHL-Pac-Sap_R"
                     /note="color: ?; sequence: 
                     CTGGTTTgaagagcttaattaagtctccttctcttaggtggcag"
     primer_bind     complement(2822..2837)
                     /label=kn2528
                     /note="SHL-3750_F2"
                     /note="color: ?; sequence: 
                     cgatgacgataagtctaggctcgagtaggaggtctactatgcctccacctgctctgc"
     primer_bind     2847..2881
                     /label=kn527
                     /note="Gal1P_mcs_F"
                     /note="color: ?; sequence: 
                     ggtcgactagtttcaaaaattcttactttttttttggatg"
     primer_bind     complement(2847..2878)
                     /label=Partial_KN576
                     /note="color: ?; sequence: 
                     ccaaaaaaaaagtaagaatttttgaaactagt"
     promoter        2852..3516
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
                     /note="color: #ffffff; direction: BOTH"
     primer_bind     2878..2898
                     /label=kn1427
                     /note="Galsplit_F"
                     /note="color: ?; sequence: 
                     gtcctgctgttggcggaggcggtagcggccgctagtctagatggacgcaaagaagttta
                     a"
     primer_bind     2909..2930
                     /label=kn339
                     /note="gal1P_F"
                     /note="color: ?; sequence: ACATGGCATTACCACCATATAC"
     primer_bind     complement(2969..2990)
                     /label=kn583
                     /note="Gal10SQ_F"
                     /note="color: ?; sequence: gggctctttacatttccacaac"
     protein_bind    3068..3185
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
                     /note="color: #31849b"
     primer_bind     complement(3128..3168)
                     /label=kn403
                     /note="pMogR5_mog45_R"
                     /note="color: ?; sequence: 
                     ttAGTTAAAAAGGCACATCTTTTTTTCAAAAACGCGACCGGTGAAGACGAG"
     primer_bind     3159..3211
                     /label=kn404
                     /note="pMogR5_mog45_F"
                     /note="color: ?; sequence: 
                     aTGAAAAAAAGATGTGCCTTTTTAACTAAAAACAATAAAGATTCTACAATACTAGC"
     primer_bind     3290..3489
                     /label=kn335
                     /note="gal1-MogR-rec_F"
                     /note="color: ?; sequence: 
                     ATAGGATGATAATGCGATTAGTTTTTTAGCCTTTTTTTTAAAAAAATTAATCAGCGAAG
                     CGATGATTTTTTATAAAAAAATTTTTATATAAAATGAAAAGCTGCATAACCACTTTAAC
                     TAATACTTTCAACATTTTCAGTTTGTATTACTTCTTATTCAAATGTCATAAAAGTATCA
                     ACAAAAAATTGTTAATATACCTC"
     primer_bind     3383..3407
                     /label=kn601
                     /note="Gal1p100upF"
                     /note="color: ?; sequence: gcaaaaactgcataaccactttaac"
     primer_bind     3465..3489
                     /label=kn542
                     /note="Gal1pEndSQ_F"
                     /note="color: ?; sequence: caacaaaaaattgttaatatacctc"
     primer_bind     complement(3486..3516)
                     /label=kn629
                     /note="M-ST1nCas9-Gal1p_R"
                     /note="color: ?; sequence: 
                     ggtggcaagggttcgatcctctagtttacgcgttttttctccttgacgttaaagtatag
                     agg"
     primer_bind     complement(3494..3518)
                     /label=kn773
                     /note="Gal1-AutoOff-Cas9_R"
                     /note="color: ?; sequence: 
                     gcccaatggagtacttcttgtccatAGATTGTGTTAGTTTAtggttttttctccttgac
                     gttaaag"
     primer_bind     complement(3494..3518)
                     /label=kn772
                     /note="Gal1-noCRp_Cas9_R"
                     /note="color: ?; sequence: 
                     ccaatggagtacttcttgtccatggAAGTAGATTGTGTTAGTTTAtggttttttctcct
                     tgacgttaaag"
     primer_bind     complement(3494..3517)
                     /label=kn516
                     /note="Gal1p_mcs_R"
                     /note="color: ?; sequence: 
                     tccttatccggagcctctagattcgaaacccatgggttttttctccttgacgttaaag"
     primer_bind     complement(3494..3516)
                     /label=hm850
                     /note="GAL1promoter 3' btm,23"
                     /note="color: ?; sequence: gttttttctccttgacgttaaag"
     primer_bind     complement(3494..3516)
                     /label=kn1428
                     /note="Galsplit_R"
                     /note="color: ?; sequence: 
                     ATCCTCCTCCGCCTGAACCTCCACCGCCgacgtccatgttttttctccttgacgttaaa
                     g"
     primer_bind     3499..3546
                     /label=al008
                     /note="NLS_CDA1(Nter_27aa_Cut)_RE:NcoI,51"
                     /note="color: ?; sequence: 
                     acgtcaaggagaaaaaaccGCTCTTCgatgtccgtgtcgcatagatgctac"
     primer_bind     complement(3500..3516)
                     /label=kn405
                     /note="LacZ-pGal_R"
                     /note="color: ?; sequence: 
                     GACGGCCAGTGAATCCGTAATCATGGTCATTATAGTTTTTTCTCCTTGACG"
     primer_bind     complement(3500..3516)
                     /label=kn338
                     /note="color: ?; sequence: TATAGTTTTTTCTCCTTGACG"
     primer_bind     complement(3501..3528)
                     /label=al039
                     /note="color: ?; sequence: cGAAGAGCgctggttttttctccttgac"
     primer_bind     complement(3503..3517)
                     /label=kn1720
                     /note="P2A-pGal_R"
                     /note="color: ?; sequence: 
                     ttccacatctccagcctgcttcagcaggctgaagttagtagcgccggctaccatgggtt
                     ttttctccttg"
     primer_bind     3509..3546
                     /label=al036
                     /note="color: ?; sequence: 
                     gaaaaaaccagcGCTCTTCgatgtcgcatagatgctac"
     regulatory      3517
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     regulatory      3520
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     primer_bind     3531..3547
                     /label=al083
                     /note="V46,46"
                     /note="color: ?; sequence: 
                     gattacaaagacgatgacgataagtctaggtcgcatagatgctacg"
     CDS             3532..3894
                     /codon_start=1
                     /label=PmCDA1
                     /note="color: #00ccff"
                     /translation="SHRCYVLFELKRRGERRACFWGYAVNKPQSGTERGIHAEIFSIRK
                     VEEYLRDNPGQFTINWYSSWSPCADCAEKILEWYNQELRGNGHTLKIEACKLYYEKNAR
                     NQIGLQNLRDNGVGLNV"
     primer_bind     3532..3566
                     /label=al064
                     /note="W122E Y34C,35"
                     /note="color: ?; sequence: 
                     tcgcatagatgctgcgttctctttgaattaaaacg"
     primer_bind     3532..3547
                     /label=al086
                     /note="V47,46"
                     /note="color: ?; sequence: 
                     TTTTTCAAGACCGAGATTACACTGGCCAATtcgcatagatgctacg"
     primer_bind     complement(3535..3567)
                     /label=al063
                     /note="W122E L36C,33"
                     /note="color: ?; sequence: 
                     tcgttttaattcaaagcaaacgtagcatctatg"
     CDS             3538..3540
                     /label=substrate binding?
                     /note="color: #993366; direction: RIGHT"
     primer_bind     3539..3571
                     /label=al062
                     /note="W122E L36C,33"
                     /note="color: ?; sequence: 
                     gatgctacgtttgctttgaattaaaacgacggg"
     unsure          3544..3546
                     /label=Expose Y to C
                     /note="color: #ff9900"
     primer_bind     complement(3546..3599)
                     /label=al087
                     /note="color: ?; sequence: 
                     tagccccaaaaacatcgttttaattcaaagagaacg"
     unsure          3550..3552
                     /label=Expose L to C
                     /note="color: #ff9900"
     primer_bind     3552..3604
                     /label=al088
                     /note="color: ?; sequence: 
                     ctttgaattaaaacgatgtttttggggctatgctg"
     primer_bind     complement(3572..3604)
                     /label=al076
                     /note="W122E W139R F49D,33"
                     /note="color: ?; sequence: 
                     cagcatagccccaatcacacgctctacgttcac"
     primer_bind     complement(3577..3601)
                     /label=al061
                     /note="W122E W50E,25"
                     /note="color: ?; sequence: catagccttcaaaacacgctctacg"
     primer_bind     complement(3583..3608)
                     /label=al051
                     /note="Y52Q,26"
                     /note="color: ?; sequence: ttcacagcttggccccaaaaacacgc"
     primer_bind     3584..3618
                     /label=al075
                     /note="W122E W139R Y52N,35"
                     /note="color: ?; sequence: 
                     cgtgtttttggggcaatgctgtgaataaaccacag"
     unsure          3589..3591
                     /label=Expose F to D
                     /note="color: #00ffff"
     unsure          3592..3594
                     /label=Expose W to E
                     /note="color: #cc99ff"
     unsure          3598..3600
                     /label=Expose Y to N, could try one more time
                     /note="color: #00ffff"
     primer_bind     complement(3604..3623)
                     /label=kn2163
                     /note="PmCDA1_160_R"
                     /note="color: ?; sequence: ccgctctgtggtttattcac"
     unsure          3604..3606
                     /label=Expose V to T
                     /note="color: #008080"
     primer_bind     complement(3621..3650)
                     /label=al059
                     /note="W122E I65D,30"
                     /note="color: ?; sequence: atttcggcgtgatcGccacgttctgtcccg"
     primer_bind     3622..3637
                     /label=kn2162
                     /note="PmCDA1_200_F"
                     /note="color: ?; sequence: 
                     gcataaggaggcaaaaatggggacagaacgtggCa"
     primer_bind     3627..3656
                     /label=al058
                     /note="W122E I65D,30"
                     /note="color: ?; sequence: agaacgtggCgatcacgccgaaatctttag"
     unsure          3637..3639
                     /label=Expose I to D
                     /note="color: #cc99ff"
     misc_feature    3640..3747
                     /label=Catalytic Domain
                     /note="color: #ff0000"
     primer_bind     complement(3656..3685)
                     /label=al057
                     /note="W122E Y78H,30"
                     /note="color: ?; sequence: cgcgcaggtgttcttcgacttttctaatgc"
     unsure          3676..3678
                     /label=Expose Y to H
                     /note="color: #cc99ff"
     primer_bind     complement(3697..3726)
                     /label=al049
                     /note="Y91K,30"
                     /note="color: ?; sequence: ccaggatgacttccaatttatcgtgaattg"
     primer_bind     3707..3736
                     /label=al048
                     /note="Y91K,30"
                     /note="color: ?; sequence: taaattggaagtcatcctggagtccttgtg"
     misc_structure  3712..3714
                     /label=substrate contact
                     /note="color: #a6acb3; direction: RIGHT"
     unsure          3715..3717
                     /label=Expose in Catalytic domain K
                     /note="color: #ff00ff"
     unsure          3724..3726
                     /label=Expose in Catalytic domain
                     /note="color: #993300"
     primer_bind     3729..3748
                     /label=kn711_PmCDA1-mid_F
                     /note="color: ?; sequence: tccttgtgcagattgcgctg"
     primer_bind     3730..3765
                     /label=kn859
                     /note="HdAdar-PmCDA1_C-term_F"
                     /note="color: ?; sequence: 
                     tattaatactgcaccatgtggtgactgcgctgaaaagatcttagaatgg"
     primer_bind     3745..3774
                     /label=al073
                     /note="W122E W139R L105N,30"
                     /note="color: ?; sequence: gctgaaaagatcaacgaatggtataaccag"
     unsure          3757..3759
                     /label=Expose L to N
                     /note="color: #00ffff"
     primer_bind     3766..3795
                     /label=al071
                     /note="W122E W139R L112D,30"
                     /note="color: ?; sequence: tataaccaggaggaccgggggaacggccac"
     unsure          3778..3780
                     /label=Expose L to D
                     /note="color: #00ffff"
     primer_bind     complement(3789..3823)
                     /label=al089
                     /note="color: ?; sequence: 
                     agagtttgcaagcttggattttcaaagtgtggccg"
     primer_bind     complement(3793..3820)
                     /label=al047
                     /note="color: ?; sequence: gtttgcaagcttcgattttcaaagtgtg"
     primer_bind     3794..3828
                     /label=al090
                     /note="color: ?; sequence: 
                     acactttgaaaatccaagcttgcaaactctattac"
     primer_bind     complement(3795..3832)
                     /label=al045
                     /note="color: ?; sequence: 
                     tctcgtaatagtttttgcaagcccagattttcaaagtg"
     primer_bind     3801..3828
                     /label=al046
                     /note="color: ?; sequence: gaaaatcgaagcttgcaaactctattac"
     unsure          3808..3810
                     /label=W122E
                     /note="color: #ff0000"
     primer_bind     3811..3847
                     /label=al034
                     /note="KN1252-PmCDA1 substituted based 126-128,38"
                     /note="color: ?; sequence: 
                     ggcttgcaaacgtgatgaggagaaaaatgcgaggaatc"
     primer_bind     complement(3812..3837)
                     /label=al043
                     /note="color: ?; sequence: atttttctcctgatagagtttgcaag"
     primer_bind     3813..3842
                     /label=al069
                     /note="W122E W139R Y128D,30"
                     /note="color: ?; sequence: ttgcaaactctatgacgagaaaaatgcgag"
     primer_bind     3813..3842
                     /label=al044
                     /note="color: ?; sequence: ttgcaaaaactattacgagaaaaatgcgag"
     primer_bind     complement(3816..3847)
                     /label=al080
                     /note="W122E W139R K130E,32"
                     /note="color: ?; sequence: 
                     gattcctcgcattttcctcgtaatagagtttg"
     primer_bind     3817..3840
                     /label=kn863
                     /note="HydraAdar-PmCDA1-flap_F"
                     /note="color: ?; sequence: 
                     ttgcatgggattgttgttggtaaactctattacgagaaaaatgcg"
     primer_bind     3818..3847
                     /label=al081
                     /note="W122E W139R K130E,30"
                     /note="color: ?; sequence: aactctattacgaggaaaatgcgaggaatc"
     primer_bind     3818..3842
                     /label=al042
                     /note="color: ?; sequence: aactctatcaggagaaaaatgcgag"
     unsure          3820..3822
                     /label=Expsose N (V150 Y128 interaction)
                     /note="color: #ff00ff"
     primer_bind     complement(3822..3857)
                     /label=al078
                     /note="W122E W139R R133E,36"
                     /note="color: ?; sequence: 
                     agcccaatttgattctccgcatttttctcgtaatag"
     unsure          3826..3828
                     /label=Expose Q (V150 L126 interaction)
                     /note="color: #ff00ff"
     unsure          3826..3828
                     /label=Expose Y to D
                     /note="color: #00ffff"
     unsure          3829..3843
                     /label=Maybe the DNA bing site
                     /note="color: #800080"
     primer_bind     3842..3871
                     /label=al067
                     /note="color: ?; sequence: ggaatcaaattgggctgcaaaacctcagag"
     primer_bind     complement(3842..3871)
                     /label=al066
                     /note="color: ?; sequence: ctctgaggttttgcagcccaatttgattcc"
     primer_bind     complement(3842..3866)
                     /label=al053
                     /note="color: ?; sequence: aggtttctcagcccaatttgattcc"
     primer_bind     3847..3871
                     /label=al052
                     /note="color: ?; sequence: caaattgggctgagaaacctcagag"
     primer_bind     complement(3847..3870)
                     /label=kn681_PmCDA1_flap_Stop_R
                     /note="color: ?; sequence: 
                     aactctagattatctgaggttccacagcccaatttg"
     unsure          3850..3852
                     /label=Expose I to E
                     /note="color: #ff9900"
     unsure          3859..3861
                     /label=W139Q
                     /note="color: #ff0000"
     misc_feature    3868..3873
                     /label=H130A R131E
                     /note="color: #a6acb3"
     primer_bind     complement(3876..3920)
                     /label=al084
                     /note="V46 V47,65"
                     /note="color: ?; sequence: 
                     TCCGAATCTCTCCTGAACCTCCACCgacgtcgctaccgcctccgcctacattcaaccca
                     accccg"
     primer_bind     complement(3876..3891)
                     /label=al040
                     /note="color: ?; sequence: agtggtctccggtattcaacccaaccccg"
     primer_bind     complement(3877..3896)
                     /label=al037
                     /note="color: ?; sequence: agtggtctcctacattcaacccaacccc"
     primer_bind     3891..3935
                     /label=al038
                     /note="30-150,45"
                     /note="color: ?; sequence: 
                     tgtaGGCGGaGGtGGcAGTGGCGGgGGTGGCTCGGGtGGgGGaGG"
     unsure          3892..3894
                     /label=Expose T (L126 Y128 interaction)
                     /note="color: #ff00ff"
     primer_bind     3894..3935
                     /label=al025
                     /note="Annealing GS linker,45"
                     /note="color: ?; sequence: 
                     gaaaGGCGGaGGtGGcAGTGGCGGgGGTGGCTCGGGtGGgGGaGG"
     CDS             3895..3984
                     /codon_start=1
                     /label=6xGS Linker
                     /note="color: #ccffcc"
                     /translation="GGGGSGGGGSGGGGSGGGGSGGGGSGGGGS"
     primer_bind     complement(3895..3939)
                     /label=hm248
                     /note="G4S btm c,45"
                     /note="color: ?; sequence: 
                     actgCCtCCcCCaCCCGAGCCACCcCCGCCACTgCCaCCtCCGCC"
     primer_bind     complement(3895..3936)
                     /label=hm244
                     /note="G4S btm a,45"
                     /note="color: ?; sequence: 
                     cgaGCCtCCcCCaCCCGAGCCACCcCCGCCACTgCCaCCtCCGCC"
     primer_bind     3895..3935
                     /label=al041
                     /note="V150T,45"
                     /note="color: ?; sequence: 
                     taccGGCGGaGGtGGcAGTGGCGGgGGTGGCTCGGGtGGgGGaGG"
     primer_bind     complement(3895..3935)
                     /label=al027
                     /note="Annealing GS linker,48"
                     /note="color: ?; sequence: 
                     catgGAtCCtCCcCCaCCCGAGCCACCcCCGCCACTgCCaCCtCCGCC"
     primer_bind     complement(3895..3935)
                     /label=hm250
                     /note="G4S btm BamHI,44"
                     /note="color: ?; sequence: 
                     gatcCtCCcCCaCCCGAGCCACCcCCGCCACTgCCaCCtCCGCC"
     primer_bind     3895..3935
                     /label=hm247
                     /note="G4S top b,45"
                     /note="color: ?; sequence: 
                     gtccGGCGGaGGtGGcAGTGGCGGgGGTGGCTCGGGtGGgGGaGG"
     primer_bind     complement(3895..3935)
                     /label=hm246
                     /note="G4S btm b,45"
                     /note="color: ?; sequence: 
                     ggacCCtCCcCCaCCCGAGCCACCcCCGCCACTgCCaCCtCCGCC"
     primer_bind     3895..3935
                     /label=hm245
                     /note="G4S top a,45"
                     /note="color: ?; sequence: 
                     ctcgGGCGGaGGtGGcAGTGGCGGgGGTGGCTCGGGtGGgGGaGG"
     primer_bind     3895..3935
                     /label=hm243
                     /note="G4S top fok,45"
                     /note="color: ?; sequence: 
                     tttcGGCGGaGGtGGcAGTGGCGGgGGTGGCTCGGGtGGgGGaGG"
     primer_bind     3895..3934
                     /label=hm249
                     /note="G4S top c,44"
                     /note="color: ?; sequence: 
                     cagtGGCGGaGGtGGcAGTGGCGGgGGTGGCTCGGGtGGgGGaG"
     primer_bind     3936..3983
                     /label=al026
                     /note="Annealing GS linker,48"
                     /note="color: ?; sequence: 
                     cagtGGCGGaGGtGGcAGTGGCGGgGGTGGCTCGGGtGGgGGaGGaTC"
     primer_bind     3936..3979
                     /label=hm249
                     /note="G4S top c,44"
                     /note="color: ?; sequence: 
                     cagtGGCGGaGGtGGcAGTGGCGGgGGTGGCTCGGGtGGgGGaG"
     primer_bind     3940..4002
                     /label=al018
                     /note="color: ?; sequence: 
                     catggGGCGGaGGtGGcAGTacgcgtatggacaagaagTACTCC"
     primer_bind     complement(3940..3987)
                     /label=al027
                     /note="Annealing GS linker,48"
                     /note="color: ?; sequence: 
                     catgGAtCCtCCcCCaCCCGAGCCACCcCCGCCACTgCCaCCtCCGCC"
     primer_bind     complement(3940..3983)
                     /label=hm250
                     /note="G4S btm BamHI,44"
                     /note="color: ?; sequence: 
                     gatcCtCCcCCaCCCGAGCCACCcCCGCCACTgCCaCCtCCGCC"
     primer_bind     3940..3980
                     /label=al041
                     /note="V150T,45"
                     /note="color: ?; sequence: 
                     taccGGCGGaGGtGGcAGTGGCGGgGGTGGCTCGGGtGGgGGaGG"
     primer_bind     3940..3980
                     /label=al025
                     /note="Annealing GS linker,45"
                     /note="color: ?; sequence: 
                     gaaaGGCGGaGGtGGcAGTGGCGGgGGTGGCTCGGGtGGgGGaGG"
     primer_bind     3940..3980
                     /label=al038
                     /note="30-150,45"
                     /note="color: ?; sequence: 
                     tgtaGGCGGaGGtGGcAGTGGCGGgGGTGGCTCGGGtGGgGGaGG"
     primer_bind     complement(3940..3980)
                     /label=hm248
                     /note="G4S btm c,45"
                     /note="color: ?; sequence: 
                     actgCCtCCcCCaCCCGAGCCACCcCCGCCACTgCCaCCtCCGCC"
     primer_bind     3940..3980
                     /label=hm247
                     /note="G4S top b,45"
                     /note="color: ?; sequence: 
                     gtccGGCGGaGGtGGcAGTGGCGGgGGTGGCTCGGGtGGgGGaGG"
     primer_bind     complement(3940..3980)
                     /label=hm246
                     /note="G4S btm b,45"
                     /note="color: ?; sequence: 
                     ggacCCtCCcCCaCCCGAGCCACCcCCGCCACTgCCaCCtCCGCC"
     primer_bind     3940..3980
                     /label=hm245
                     /note="G4S top a,45"
                     /note="color: ?; sequence: 
                     ctcgGGCGGaGGtGGcAGTGGCGGgGGTGGCTCGGGtGGgGGaGG"
     primer_bind     complement(3940..3980)
                     /label=hm244
                     /note="G4S btm a,45"
                     /note="color: ?; sequence: 
                     cgaGCCtCCcCCaCCCGAGCCACCcCCGCCACTgCCaCCtCCGCC"
     primer_bind     3940..3980
                     /label=hm243
                     /note="G4S top fok,45"
                     /note="color: ?; sequence: 
                     tttcGGCGGaGGtGGcAGTGGCGGgGGTGGCTCGGGtGGgGGaGG"
     primer_bind     3982..4002
                     /label=al020
                     /note="color: ?; sequence: 
                     CCGGGTACTTCAGAATCCGCTACGCCGGAGtcCatggacaagaagTACTCC"
     primer_bind     3983..4012
                     /label=kn559_Cas9_Nco1_F
                     /note="color: ?; sequence: 
                     aaacccATGGACAAGAAGTACTCCATTGGGCTCG"
     primer_bind     3983..4012
                     /label=kn559
                     /note="color: ?; sequence: 
                     aaacccATGGACAAGAAGTACTCCATTGGGCTCG"
     primer_bind     3983..4010
                     /label=kn947_Cas9_nco_F
                     /note="color: ?; sequence: ccatggacaagaagtactccattgggct"
     primer_bind     3983..4009
                     /label=hm042
                     /note="NcoI-Cas9 5,42"
                     /note="color: ?; sequence: 
                     TAAGAAGGAGATATACCATGgacaagaagtactccattgggc"
     primer_bind     3983..4002
                     /label=kn1625
                     /note="OR1-Apa-Cas_F"
                     /note="color: ?; sequence: 
                     ggtgataatggttgcagggcccATGGACAAGAAGTACTCC"
     CDS             3985..8088
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /note="This feature has 3 segments:
                        1: 3985 .. 4011 / #ff99cc
                        2: 4012 .. 4014 / #ff6600
                        3: 4015 .. 8088 / #ff99cc"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     3985..4002
                     /label=al016
                     /note="color: ?; sequence: 
                     gaaggaagccgctgcaaaagaggccgcagctaaagctatggacaagaagTACTCC"
     misc_feature    3985..3986
                     /label=truncated 161
                     /note="color: #a6acb3"
     primer_bind     3988..4008
                     /label=kn2565
                     /note="Cas9_Nhe_F"
                     /note="color: ?; sequence: 
                     cctggcgctagcggtGACAAGAAGTACTCCATTGGG"
     primer_bind     complement(3990..4012)
                     /label=kn982
                     /note="Cas9_30_R"
                     /note="color: ?; sequence: cgagcccaatggagtacttcttg"
     primer_bind     3991..4028
                     /label=kn1499
                     /note="Cas9_D10_F"
                     /note="color: ?; sequence: 
                     aagaagtactccattgggctcgAtatcggcacaaacag"
     primer_bind     complement(3996..4031)
                     /label=kn553_Cas9_D10A_R
                     /note="color: ?; sequence: 
                     ACGCTGTTTGTGCCGATAgCGAGCCCAATGGAGTAC"
     primer_bind     3997..4032
                     /label=kn552_Cas9_D10A_F
                     /note="color: ?; sequence: 
                     TACTCCATTGGGCTCGcTATCGGCACAAACAGCGTC"
     primer_bind     4012..4032
                     /label=kn652eCas9_D10A_F
                     /note="color: ?; sequence: 
                     TACTCAATAGGCTTAGcTATCGGCACAAATAGCGTC"
     primer_bind     4066..4089
                     /label=kn2564
                     /note="Cas9-90_Nhe_F"
                     /note="color: ?; sequence: 
                     cctggcgctagcggtccgAGCAAAAAATTCAAAGTTCTG"
     primer_bind     complement(4080..4103)
                     /label=hm511
                     /note="Cas9 30th aa reverse for seq,24"
                     /note="color: ?; sequence: cgatcggtattgcccagaactttg"
     primer_bind     complement(4080..4099)
                     /label=kn771_Cas9_100_R
                     /note="color: ?; sequence: cggtattgcccagaactttg"
     primer_bind     4091..4111
                     /label=kn1388
                     /note="Cas9_150_F"
                     /note="color: ?; sequence: GCAATACCGATCGCCACAGCA"
     primer_bind     4120..4136
                     /label=kn2563
                     /note="Cas9-120_Nhe_F"
                     /note="color: ?; sequence: 
                     cctggcgctagcggtAACCTCATTGGCGCCCT"
     primer_bind     complement(4152..4181)
                     /label=TA321 2A-Cas-2
                     /note="color: ?; sequence: cttttgagccgcgtggcttcggccgtttcc"
     primer_bind     complement(4162..4187)
                     /label=hm129
                     /note="cas9 a btm ,35"
                     /note="color: ?; sequence: 
                     ctgggtctcgctgttcttttgagccgcgtggcttc"
     primer_bind     4182..4208
                     /label=hm130
                     /note="cas9 b top ,45"
                     /note="color: ?; sequence: 
                     gctggtctcaacagcacggcgcagatatacccgcag"
     primer_bind     4211..4230
                     /label=kn1014_Cas9_250_F
                     /note="color: ?; sequence: AGAATCGGATCTGCTACCTG"
     primer_bind     complement(4244..4264)
                     /label=kn1010_Cas9_250_R
                     /note="color: ?; sequence: CCACCTTAGCCATCTCATTAC"
     primer_bind     4415..4434
                     /label=kn1402
                     /note="Cas9_450_F"
                     /note="color: ?; sequence: ACAGTACTGATAAGGCTGAC"
     primer_bind     complement(4421..4439)
                     /label=kn1500
                     /note="Cas9-500_R"
                     /note="color: ?; sequence: cgcaagtcagccttatcag"
     primer_bind     complement(4443..4485)
                     /label=hm201
                     /note="Cas9 5' NdeI btm,43"
                     /note="color: ?; sequence: 
                     gtgtccccgaaatttgatcatatgcgccagcgcgagatagatc"
     primer_bind     complement(4446..4470)
                     /label=hm131
                     /note="cas9 b btm NdeI,25"
                     /note="color: ?; sequence: gatcatatgcgccagcgcgagatag"
     primer_bind     complement(4481..4499)
                     /label=kn2562
                     /note="Cas9_550_R"
                     /note="color: ?; sequence: CCCTCGATGAGGAAGTGTC"
     primer_bind     complement(4487..4506)
                     /label=kn960_Cas9-trunc_R
                     /note="color: ?; sequence: 
                     accgcggccgctacccaggtccccctcgatgagga"
     primer_bind     complement(4527..4546)
                     /label=hm570
                     /note="Cas9_0.5k_R,20"
                     /note="color: ?; sequence: gttggataaagagtttgtcg"
     primer_bind     complement(4578..4598)
                     /label=kn1369
                     /note="Cas9_Split204_R"
                     /note="color: ?; sequence: 
                     TGAACCTCCACCGCCgacgtcgctagcTCCTCCACCGGATGCGTTGATCGGGTTC"
     primer_bind     4593..4615
                     /label=kn1370
                     /note="Cas9Split205_F"
                     /note="color: ?; sequence: 
                     tagcgacgtcGGCGGTGGAGGTTCAGGCGGAGGAGGATCAGGAGTTGACGCCAAAGCAA
                     "
     primer_bind     4896..4920
                     /label=kn961_Cas9_trunc_F
                     /note="color: ?; sequence: 
                     ggaggtagcggccgcggtattctgcgagtgaacacggagatc"
     primer_bind     4901..4920
                     /label=kn561_Cas9_1k_F
                     /note="color: ?; sequence: TGCGAGTGAACACGGAGATC"
     primer_bind     complement(4977..4998)
                     /label=kn560_Cas9_1k_R
                     /note="color: ?; sequence: AAGGGCCTTCAGCAAAGTCAAG"
     primer_bind     5057..5067
                     /label=hm099
                     /note="XTEN tag top,43,,"
                     /note="color: ?; sequence: 
                     ccGGaTCGGAGACGCCGGGTACTTCAGAATCCGCTACGCCGGA"
     primer_bind     complement(5166..5185)
                     /label=kn1011_Cas9_1.2K_R
                     /note="color: ?; sequence: TGCGCAACAGATCTTCTCTg"
     primer_bind     5172..5191
                     /label=hm572
                     /note="Cas9_1.2k_F,20"
                     /note="color: ?; sequence: agatctgttgcgcaaacagc"
     primer_bind     complement(5178..5188)
                     /label=kn185
                     /note="BLF1_SNP_R"
                     /note="color: ?; sequence: 
                     ctgcagcggccgctactagtGCCAGTTAATAGTTTGCGCaac"
     primer_bind     5329..5351
                     /label=kn892_yCas9XmaKO2_F
                     /note="color: ?; sequence: ccctactatgtaggccccctcgc"
     primer_bind     5337..5367
                     /label=kn882_yCas9_Xma1KO_F
                     /note="color: ?; sequence: tgtaggccccctcgcTcggggaaattccaga"
     primer_bind     complement(5571..5589)
                     /label=kn1367
                     /note="Cas9_Split535_R"
                     /note="color: ?; sequence: 
                     tGAACCTCCACCGCCgacgtcgctagcTCCTCCACCTCTCATCCCTTCTGTGACG"
     primer_bind     5589..5608
                     /label=kn1368
                     /note="Cas9_Split536_F"
                     /note="color: ?; sequence: 
                     tagcgacgtcGGCGGTGGAGGTTCaGGCGGAGGAGGATCAAAGCCAGCATTCCTGTCTG
                     "
     primer_bind     5663..5701
                     /label=kn2310
                     /note="hCas9-BbsKO_F"
                     /note="color: ?; sequence: 
                     ccgtgaaacagctcaaagaagattatttcaaaaagattg"
     primer_bind     5830..5849
                     /label=kn563_Cas9_2k_F
                     /note="color: ?; sequence: CTTGAGGACATTGTCCTCAC"
     primer_bind     5902..5923
                     /label=al003a
                     /note="dCas9_middle,22"
                     /note="color: ?; sequence: gctcatctcttcgacgacaaag"
     primer_bind     complement(5902..5923)
                     /label=kn562_Cas9_2k_R
                     /note="color: ?; sequence: CTTTGTCGTCGAAGAGATGAGC"
     primer_bind     5926..5945
                     /label=hm573
                     /note="Cas9_1.9k_F,20"
                     /note="color: ?; sequence: atgaaacagctcaagaggcg"
     primer_bind     5984..6015
                     /label=hm124
                     /note="color: ?; sequence: 
                     tcaatgggatccgagacaagcagagtggaaag"
     primer_bind     complement(6031..6051)
                     /label=kn1013_Cas9_2.1K_R
                     /note="color: ?; sequence: GGCAAATCCATCGGACTTAAG"
     primer_bind     6040..6076
                     /label=hm566
                     /note="Cas9 R691A 3' top,37"
                     /note="color: ?; sequence: 
                     GATGGATTTGCCAACGCGAACTTCATGCAGTTGATCC"
     primer_bind     complement(6040..6076)
                     /label=hm565
                     /note="Cas9 R691A 5' btm,37"
                     /note="color: ?; sequence: 
                     GGATCAACTGCATGAAGTTCGCGTTGGCAAATCCATC"
     primer_bind     6282..6301
                     /label=kn564_Casa9_2.5k_F
                     /note="color: ?; sequence: GAACCAAACTACCCAGAAGG"
     primer_bind     complement(6483..6517)
                     /label=kn550_Cas9_H840A_R
                     /note="color: ?; sequence: 
                     ACTGGGGCACGATAgcATCCACGTCGTAGTCGGAG"
     primer_bind     6484..6518
                     /label=kn551_Cas9_H840A_F
                     /note="color: ?; sequence: 
                     TCCGACTACGACGTGGATgcTATCGTGCCCCAGTC"
     primer_bind     complement(6555..6586)
                     /label=kn731_yCas9_N863A_R
                     /note="color: ?; sequence: 
                     CACTCTTCCCTCTAgcTTTATCGGATCTTGTC"
     primer_bind     6557..6587
                     /label=kn730_yCas9_N863A_F
                     /note="color: ?; sequence: CAAGATCCGATAAAgcTAGAGGGAAGAGTGA"
     primer_bind     6576..6603
                     /label=sy210
                     /note="tale_nCas9_Fw"
                     /note="color: ?; sequence: AGGGAAGAGTGATAACGTCCCCTCAGAA"
     primer_bind     6643..6664
                     /label=hm513
                     /note="Cas9 887th aa fw for seq,22"
                     /note="color: ?; sequence: ctgaacgccaaactgatcacac"
     primer_bind     6675..6694
                     /label=hm574
                     /note="Cas9_2.7k_F,20"
                     /note="color: ?; sequence: cgataatctgactaaggctg"
     primer_bind     complement(7071..7098)
                     /label=sy209
                     /note="tale_nCas9_Rv"
                     /note="color: ?; sequence: AAAGAAGTACTTAGCGGTGGCCTTGCCT"
     primer_bind     complement(7122..7146)
                     /label=al085
                     /note="V47,25"
                     /note="color: ?; sequence: ATTGGCCAGTGTAATCTCGGTCTTG"
     primer_bind     7196..7242
                     /label=al091
                     /note="color: ?; sequence: 
                     aaatcgtgtgggacaagggtagggatttcgcgacagtccggaaggtc"
     primer_bind     7226..7265
                     /label=TA329 hCas-dAarI-1
                     /note="color: ?; sequence: 
                     cgacagtccggaaggtcctgtccatgcctcaggtgaacat"
     primer_bind     7245..7254
                     /label=TA313 GFP-con-2
                     /note="color: ?; sequence: 
                     tgtttaattaaTTACTTGTACAGCTCGTCCATGCCG"
     primer_bind     7245..7254
                     /label=TA877 R9-GFP-2
                     /note="color: ?; sequence: 
                     AGCTCGAATTCGGATCCTCTAGCTTGTACAGCTCGTCCATGCCG"
     primer_bind     7292..7310
                     /label=kn1047_Cas9_3.3k_F
                     /note="color: ?; sequence: gaggcttctccaaggaaag"
     primer_bind     7390..7409
                     /label=kn566_Cas9_3.5k_F
                     /note="color: ?; sequence: TCTCCTACAGTCGCTTACAG"
     primer_bind     7421..7440
                     /label=hm575
                     /note="Cas9_3.4k_F,20"
                     /note="color: ?; sequence: tggccaaagtggagaaaggg"
     primer_bind     complement(7468..7487)
                     /label=kn565_Cas9_3.5k_R
                     /note="color: ?; sequence: ATTGTGATGCCCAGCAGTTC"
     primer_bind     7502..7521
                     /label=kn1477
                     /note="Cas9_3.5k_F"
                     /note="color: ?; sequence: GCTTCGAAAAAAACCCCATC"
     primer_bind     7522..7552
                     /label=hm054
                     /note="XhoI Cas9 3.5k-2,31"
                     /note="color: ?; sequence: gactttctcgaggcgaaaggatataaagagg"
     primer_bind     complement(7642..7663)
                     /label=kn1389
                     /note="Cas9_3.5k_R"
                     /note="color: ?; sequence: CCAGCTCGTTACCTTTCTGCAG"
     primer_bind     complement(7730..7751)
                     /label=kn914_Cas9_3.7K_R
                     /note="color: ?; sequence: tgcttctgctcattatcttcgg"
     primer_bind     complement(7824..7841)
                     /label=kn1063_hCas9_4k_R
                     /note="color: ?; sequence: attagcgtcggccaggatc"
     primer_bind     complement(7888..7909)
                     /label=sy262
                     /note="color: ?; sequence: tgttttctgcctgctccctgat"
     primer_bind     7888..7909
                     /label=sy261
                     /note="color: ?; sequence: atcagggagcaggcagaaaaca"
     primer_bind     7891..7908
                     /label=kn2575
                     /note="Cas9-3.9k_F"
                     /note="color: ?; sequence: agggagcaggcagaaaac"
     primer_bind     7896..7917
                     /label=hm132
                     /note="Cas9 AscI top,22"
                     /note="color: ?; sequence: gcaggcagaaaacattatccac"
     primer_bind     7916..7945
                     /label=TA148 hCas-9
                     /note="color: ?; sequence: acttgtttactctgaccaacttgggcgcgc"
     primer_bind     7921..7962
                     /label=al021
                     /note="Cut to 161 site of KN1252,42"
                     /note="color: ?; sequence: 
                     tttactctgaccaacttgggcgcgcctgcagccttcaagtac"
     primer_bind     complement(7945..7969)
                     /label=TA331 AID-dAarI-2
                     /note="color: ?; sequence: tgtcgaagtacttgaaggctgcagg"
     primer_bind     8041..8060
                     /label=kn725_Cas9_4k_F
                     /note="color: ?; sequence: ACGGGGCTCTATGAAACAAG"
     primer_bind     complement(8070..8088)
                     /label=hm055
                     /note="Cas9 stop 3'XhoI,48"
                     /note="color: ?; sequence: 
                     gggctcgaggtaccgcggctagcttactagtctccaccgagctgagag"
     primer_bind     complement(8071..8088)
                     /label=kn1624
                     /note="LVA-Cas9_R"
                     /note="color: ?; sequence: 
                     GCCGttatgcaaccagtctagaGTCTCCACCGAGCTGAGA"
     primer_bind     8079..8145
                     /label=kn1497
                     /note="OR1-Ugi_F"
                     /note="color: ?; sequence: 
                     ctattttacctctggcggtgataatggttgcagggcccATGACCAACCTTTCCGAC"
     primer_bind     complement(8096..8121)
                     /label=kn923
                     /note="attL2-Cas9_R"
                     /note="color: ?; sequence: 
                     ccaactttgtacaagaaagctgggttaTCACACCTTCCTCTTCTTCTTGGGGTCAG"
     primer_bind     complement(8098..8124)
                     /label=kn556_GSlink-NLS-Cas9_R
                     /note="color: ?; sequence: 
                     CCTCCACCTCCaGAACCTCCTCCACCCACCTTCCTCTTCTTCTTGGGGTC"
     primer_bind     complement(8100..8121)
                     /label=hm125
                     /note="Cas9 term btm NotI"
                     /note="color: ?; sequence: 
                     ggggcggccgcttatcacaccttcctcttcttcttgggg"
     primer_bind     complement(8100..8120)
                     /label=kn593_NLS-Cas9_R
                     /note="color: ?; sequence: 
                     gcccaatggagtacttcttgtccatgaccttccgcttcttctttggg"
     primer_bind     complement(8101..8135)
                     /label=al092
                     /note="color: ?; sequence: 
                     TTGGTCATggtacccaccttcctcttcttcttggg"
     primer_bind     8101..8123
                     /label=kn1085_ApaNLS_F
                     /note="color: ?; sequence: 
                     TgGAGGaGGGcccaagaagaagaggaaggtggG"
     primer_bind     8101..8122
                     /label=kn1296
                     /note="Gal10-spe-NLS_F"
                     /note="color: ?; sequence: 
                     aaaagtaagaatttttgaaactagtatgCCCAAGAAGAAGAGGAAGGTGG"
     CDS             8101..8121
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     primer_bind     complement(8101..8121)
                     /label=hm498
                     /note="Cas9 NLS 3' btm w SphI,35"
                     /note="color: ?; sequence: 
                     ttaGCATGCtatcacaccttcctcttcttcttggg"
     primer_bind     complement(8101..8121)
                     /label=hm567
                     /note="Cas9 R691A 3' btm,45"
                     /note="color: ?; sequence: 
                     CGAATTCGGATCCTCTAGTTATCACACCTTCCTCTTCTTCTTGGG"
     primer_bind     complement(8101..8121)
                     /label=al003
                     /note="dCas9_XbaI,45"
                     /note="color: ?; sequence: 
                     tccttatccggagcctctagactacaccttcctcttcttcttggg"
     primer_bind     complement(8101..8121)
                     /label=kn946_Cas9-nosT_R
                     /note="color: ?; sequence: 
                     tcgagctccctaactagtcaccttcctcttcttcttggg"
     primer_bind     8101..8120
                     /label=kn588_Flag-NLS_F
                     /note="color: ?; sequence: 
                     tgattacaaagacgatgacgataagtctagtCCCAAGAAGAAGAGGAAGGT"
     primer_bind     8101..8119
                     /label=kn1266
                     /note="Gal10-Spe-NLS_F"
                     /note="color: ?; sequence: 
                     gcgtccatccaaaaaaaaagtaagaatttttgaaactagtatgCCCAAGAAGAAGAGGA
                     AGG"
     primer_bind     complement(8102..8123)
                     /label=kn1476
                     /note="Xho-Xba-Apa-NLS_R"
                     /note="color: ?; sequence: 
                     GACATAACTAATTACATGACTCGAGtctagaaccgggcccCACCTTCCTCTTCTTCTTG
                     G"
     primer_bind     complement(8102..8121)
                     /label=Sy424
                     /note="GS linker SY20"
                     /note="color: ?; sequence: 
                     CCtCCGCCgaaattcaccttcctcttcttcttgg"
     primer_bind     complement(8102..8121)
                     /label=sy317
                     /note="color: ?; sequence: caccttcctcttcttcttgg"
     primer_bind     complement(8103..8121)
                     /label=hm661
                     /note="Cas9-NLS_3' btm for GA w SUMOstar,44"
                     /note="color: ?; sequence: 
                     AAGGACGCGGTACCGCGGCCGCtcacaccttcctcttcttcttg"
     primer_bind     complement(8106..8145)
                     /label=kn2574
                     /note="NLS-Xba_R"
                     /note="color: ?; sequence: 
                     gtaGGAAAGGCAacctctagaacccaccttcctcttcttc"
     primer_bind     8106..8137
                     /label=al093
                     /note="color: ?; sequence: 
                     gaagaagaggaaggtgggtaccATGACCAACC"
     primer_bind     complement(8106..8125)
                     /label=kn521_T7pol-NLS_R
                     /note="color: ?; sequence: 
                     gaagtcgttcttagcgatgttaatcgtgttcatACCGACCTTCCGCTTCTTC"
     primer_bind     complement(8108..8124)
                     /label=sy268
                     /note="nCas9 Rv2"
                     /note="color: ?; sequence: TCCACCcaccttcctcttct"
     primer_bind     8122..8152
                     /label=TA296 Cugi-1
                     /note="color: ?; sequence: 
                     AGTCggtaccATGACCAACCTTTCCGACATCATAG"
     primer_bind     8126..8145
                     /label=kn2052
                     /note="Ugi_NLS-Apa_F"
                     /note="color: ?; sequence: 
                     CTAAGAAGAAACGTAAAGTAgggcCcATGACCAACCTTTCCGAC"
     CDS             8128..8379
                     /codon_start=1
                     /label=Ugi
                     /note="color: #993366"
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     primer_bind     8128..8151
                     /label=TA300 ugi-2A-1
                     /note="color: ?; sequence: 
                     TATAGGGAGACCCAAGCTGGATGACCAACCTTTCCGACATCATA"
     primer_bind     8128..8147
                     /label=kn1204
                     /note="OR1-SD-Ugi_F"
                     /note="color: ?; sequence: 
                     tggcggtgataatggttgcagggccgAGGAGGGCCATCAAATGACCAACCTTTCCGACA
                     T"
     primer_bind     8128..8147
                     /label=TA257 AoUgi-1
                     /note="color: ?; sequence: 
                     ACCAACCAAACCACCCAAggATGACCAACCTTTCCGACAT"
     primer_bind     8128..8145
                     /label=kn1461
                     /note="PmCDA1-Ugi_F"
                     /note="color: ?; sequence: 
                     gtaaaaatactccacaccactaagagtcctgctgtttctagaATGACCAACCTTTCCGA
                     C"
     primer_bind     8128..8142
                     /label=kn1430_Gal10-Ugi_F
                     /note="color: ?; sequence: 
                     gcgtccatccaaaaaaaaagtaagaatttttgaaactagtATGACCAACCTTTCC"
     primer_bind     8130..8154
                     /label=kn2531
                     /note="Flag-Xho-UGI_F"
                     /note="color: ?; sequence: 
                     acgataagtctaggctcgagACgAACCTTTCCGACATCATAGAG"
     primer_bind     complement(8212..8229)
                     /label=kn2166
                     /note="UgiN1-Eag_R"
                     /note="color: ?; sequence: 
                     CGAGCTCGGTACCCGGCCGttatgcaaccagCCCAATGACCTCCTCAAC"
     primer_bind     8227..8245
                     /label=kn2164
                     /note="PmCDA1N1-UgiC1_F"
                     /note="color: ?; sequence: 
                     tgaataaaccacagagcggtggagggAATAAGCCGGAAAGTG"
     primer_bind     complement(8252..8271)
                     /label=kn1690
                     /note="LVA-Ugi_R3"
                     /note="color: ?; sequence: 
                     GTGAATTCGAGCTCGGTACCCGGCCGttatgcaaccagATCATACGCAGTGTGTACGA"
     primer_bind     complement(8290..8307)
                     /label=kn1691
                     /note="LVA-Ugi_R4"
                     /note="color: ?; sequence: 
                     GTGAATTCGAGCTCGGTACCCGGCCGttatgcaaccagTGACGTGAGCAGCATCAC"
     primer_bind     complement(8333..8352)
                     /label=kn1689
                     /note="LVA-Ugi_R2"
                     /note="color: ?; sequence: 
                     GTGAATTCGAGCTCGGTACCCGGCCGttatgcaaccagAGAGTCCTGAATCACCAGAG"
     primer_bind     complement(8355..8381)
                     /label=TA330 cugi-dAarI-2
                     /note="color: ?; sequence: 
                     GTTTAAACTTAAGCTTGCGGCCGCttaTAGCATCTTGATCTTGTTCTCTCCA"
     primer_bind     complement(8355..8381)
                     /label=TA490 cugi-trans-2
                     /note="color: ?; sequence: 
                     TAAGCTTGCGGCCGCTTTATAGCATCTTGATCTTGTTCTCTCCA"
     primer_bind     complement(8355..8379)
                     /label=TA297 Cugi-2
                     /note="color: ?; sequence: 
                     TTCCCCTGCCTTCTAGCATCTTGATCTTGTTCTCTCCA"
     primer_bind     complement(8356..8382)
                     /label=kn2532
                     /note="Xba-stop-UGI_R"
                     /note="color: ?; sequence: 
                     cggtcattctagacctcctaTAGCATCTTGATCTTGTTCTCTCC"
     primer_bind     complement(8356..8379)
                     /label=hm970
                     /note="color: ?; sequence: TAGCATCTTGATCTTGTTCTCTCC"
     primer_bind     complement(8358..8379)
                     /label=TA871 R9-Cas-ugi-2
                     /note="color: ?; sequence: 
                     AGCTCGAATTCGGATCCTCTAGTAGCATCTTGATCTTGTTCTCT"
     primer_bind     complement(8359..8379)
                     /label=kn2533
                     /note="Xba-stop-UGI-LVA_R"
                     /note="color: ?; sequence: 
                     tcattctagacctcctacgcaacTAGCATCTTGATCTTGTTCTC"
     primer_bind     complement(8359..8379)
                     /label=kn2165
                     /note="PmCDA1C1-stop-UGI_R"
                     /note="color: ?; sequence: 
                     ccatTTTTGCCTCCttatgcaacTAGCATCTTGATCTTGTTCTC"
     primer_bind     complement(8361..8380)
                     /label=hm127
                     /note="color: ?; sequence: 
                     ggggcggccgcttatcaTAGCATCTTGATCTTGTTC"
     primer_bind     complement(8361..8379)
                     /label=kn1496
                     /note="dCas-Ugi_R"
                     /note="color: ?; sequence: 
                     GAGTATTTCTTATCCATTTTTGCCTCCTgAAATgggccTAGCATCTTGATCTTGTTC"
     primer_bind     complement(8361..8379)
                     /label=kn2301
                     /note="Ugi-stop_R"
                     /note="color: ?; sequence: 
                     acgaCGGCCAGTGAATTCGAGCTCGGTACCCGGCCGttaccTAGCATCTTGATCTTGTT
                     C"
     primer_bind     complement(8361..8379)
                     /label=kn1460
                     /note="LVA-Ugi_R"
                     /note="color: ?; sequence: 
                     GTGAATTCGAGCTCGGTACCCGGCCGttatgcaaccagtccTAGCATCTTGATCTTGTT
                     C"
     primer_bind     complement(8364..8413)
                     /label=al094
                     /note="color: ?; sequence: 
                     cttattaggatccttatccggagcctctagactaTAGCATCTTGATCTTG"
     primer_bind     complement(8364..8379)
                     /label=kn1203
                     /note="dCas-Ugi_R"
                     /note="color: ?; sequence: 
                     gAGTATTTCTTATCCATTTTTGCCTCCTAAAATgggcccTAGCATCTTGATCTTG"
     primer_bind     8383..8433
                     /label=kn515
                     /note="Adh1T_mcs_F"
                     /note="color: ?; sequence: 
                     ggtttcgaatctagaggctccggataaggatcctaataagcgaatttcttatgatttat
                     g"
     primer_bind     complement(8384..8413)
                     /label=Partial kn574
                     /note="color: ?; sequence: cttattaggatccttatccggagcctctag"
     primer_bind     8398..8414
                     /label=al023
                     /note="Cut to 161 site of KN1252,30"
                     /note="color: ?; sequence: gaaataaACTAGTtaaggatcctaataagc"
     primer_bind     complement(8401..8426)
                     /label=kn589
                     /note="BamH1-ADH1T_R"
                     /note="color: ?; sequence: cataagaaattcgcttattaggatcc"
     terminator      8413..8600
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for alcohol dehydrogenase 
                     1"
                     /note="color: #ffffff"
     primer_bind     complement(8486..8508)
                     /label=kn569
                     /note="color: ?; sequence: tcgttttaaaacctaagagtcac"
     primer_bind     8551..8572
                     /label=kn1362
                     /note="adh1T_end_F"
                     /note="color: ?; sequence: ctttctcaggtatagcatgagg"
     primer_bind     complement(8580..8601)
                     /label=kn1018
                     /note="ADH1-EagKpnSac1_R"
                     /note="color: ?; sequence: 
                     AGTGAATTCGAGCTCGGTACCCGGCCGccggtagaggtgtggtcaata"
     primer_bind     complement(8613..8672)
                     /label=kn517
                     /note="T7T-adh1T_R"
                     /note="color: ?; sequence: 
                     ccttggggcctctaaacgggtcttgagggggttttttgcccgggagcgatttgcaggca
                     t"
     primer_bind     complement(8617..8636)
                     /label=al024
                     /note="Cut to 161 site of KN1252,20"
                     /note="color: ?; sequence: tgcccgggagcgatttgcag"
     primer_bind     8635..8655
                     /label=kn534
                     /note="Ade1term-Integration-T7term_R"
                     /note="color: ?; sequence: 
                     GGTCTCACTAACGTGATTTACATATACTACAAGTCGCCAGTGTAACTCCTCACTGAATA
                     caaaaaaccccctcaagaccc"
     terminator      8636..8683
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     primer_bind     8637..8661
                     /label=hm064
                     /note="hm014 gRNA templ b3,26"
                     /note="color: ?; sequence: caaaaaacccctcaagacccgtttag"
     primer_bind     8647..8667
                     /label=kn567
                     /note="T7t_F"
                     /note="color: ?; sequence: tcaagacccgtttagaggccc"
     primer_bind     complement(8655..8713)
                     /label=kn518
                     /note="T7T-mcs_R"
                     /note="color: ?; sequence: 
                     ttggagctccaccgcggaagcttggtatgcatagcataaccccttggggcctctaaacg
                     "
     primer_bind     complement(8691..8710)
                     /label=kn1329
                     /note="Leu2d-Sac12_R"
                     /note="color: ?; sequence: 
                     gcagacattagaatggtatatccttgagctccaccgcggaagctt"
     promoter        complement(8719..8737)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(8719..8737)
                     /label=hm817
                     /note="5'-FAM T7 promoter,19"
                     /note="color: ?; sequence: TAATACGACTCACTATAGG"
     primer_bind     complement(8720..8737)
                     /label=hm444
                     /note="T7 promoter,18"
                     /note="color: ?; sequence: TAATACGACTCACTATAG"
     primer_bind     8744..8763
                     /label=kn1009
                     /note="M13FR"
                     /note="color: ?; sequence: ACTGGCCGTCGTTTTACAAC"
     primer_bind     complement(8744..8760)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(8745..8765)
                     /label=kn535
                     /note="Kin3term_integration-M13_F"
                     /note="color: ?; sequence: 
                     GATATCGATAAAATTTGCTCTGAGAACATTTATACATTAATACATACGGGTATGTATGA
                     ATCacgttgtaaaacgacggccag"
     primer_bind     complement(8796..8903)
                     /label=kn383
                     /note="LacZA3_R"
                     /note="color: ?; sequence: 
                     CATCAGGCGCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGG
                     CCTCTTCGCTATTACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGGCGATTAAG"
     primer_bind     complement(8884..8901)
                     /label=kn1626
                     /note="Erm_end_F"
                     /note="color: ?; sequence: gccATTCGCCATTCAGGC"
     rep_origin      8905..9360
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     complement(9020..9039)
                     /label=hm459
                     /note="HH gRNA seq @ f1 ori btm,20"
                     /note="color: ?; sequence: GAAAGCCGGCGAACGTGGCG"
     primer_bind     complement(9114..9133)
                     /label=kn2264
                     /note="f1_F"
                     /note="color: ?; sequence: gaaccatcaccctaatcaag"
     primer_bind     9229..9254
                     /label=hm363
                     /note="f1ori for seq of sy5 puro top for seq,26"
                     /note="color: ?; sequence: CACTCAACCCTATCTCGGTCTATTCT"
     primer_bind     9308..9329
                     /label=kn183
                     /note="pRS303ver_R"
                     /note="color: ?; sequence: GATTTAACAAAAATTTAACGCG"
     primer_bind     9364..9386
                     /label=kn503
                     /note="pRS_markerout_R"
                     /note="color: ?; sequence: 
                     gtggaattctcatgtttgacagcttatcatcgatGCGGTATTTTCTCCTTACGC"
     primer_bind     complement(9383..9413)
                     /label=hm578
                     /note="PNAS no.90,90"
                     /note="color: ?; sequence: 
                     CGGGTGTCGGGGCTGGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGCACC
                     ATATGCGGTGTGAAATACCGCACAGATGCGT"
     primer_bind     complement(9383..9413)
                     /label=hm576
                     /note="SK3 pUC19,90"
                     /note="color: ?; sequence: 
                     AATTCTCATTTTACTTACCGGACGCTATTAGCAGTGGCAGATTGTACTGAGAGTGCACC
                     ATATGCGGTGTGAAATACCGCACAGATGCGT"
     primer_bind     9383..9397
                     /label=kn1197
                     /note="ade1int-Ura3_R"
                     /note="color: ?; sequence: 
                     cGAATCATAAGCATTGCTTACAAAGAATACACATACGAAATATTAACGcatctgtgcgg
                     t"
     primer_bind     9388..9407
                     /label=kn105
                     /note="pRS_Mark_SNP_R"
                     /note="color: ?; sequence: 
                     ctgcagcggccgctactagtTCTGTGCGGTATTTCACACC"
     primer_bind     9388..9407
                     /label=kn97
                     /note="pRS-Mark-XNE_R"
                     /note="color: ?; sequence: 
                     tctagaagcggccgcgaattcTCTGTGCGGTATTTCACACC"
     primer_bind     9658..9683
                     /label=kn2478
                     /note="Leu2P_F"
                     /note="color: ?; sequence: 
                     cgcggcctttttaattaactgtgggaatactcaggtatcg"
     promoter        9660..10064
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     9660..9682
                     /label=kn2437
                     /note="2u-sph-Leu_F"
                     /note="color: ?; sequence: 
                     CTTCGTTCAGGTGgcatgcaactgtgggaatactcaggtatc"
     primer_bind     complement(9696..9716)
                     /label=kn2439
                     /note="Leu2p_R"
                     /note="color: ?; sequence: tggttgctaagagattcgaac"
     primer_bind     10048..10073
                     /label=kn1328
                     /note="Leu2d_F"
                     /note="color: ?; sequence: aaggatataccattctaatgtctgcc"
     CDS             10077..11171
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /note="color: #ff7f50"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     10077..10110
                     /label=kn2270
                     /note="P2A_Leu2_F2"
                     /note="color: ?; sequence: 
                     gcaggctggagatgtggaagagaaccctggaccaatgtctgcccctaagaagatcgtcg
                     ttttgccag"
     primer_bind     complement(10306..10327)
                     /label=kn2477
                     /note="Leu250_R"
                     /note="color: ?; sequence: gtaccccatttaggaccaccca"
     primer_bind     10325..10344
                     /label=kn1332
                     /note="Leu2_300-NsiHind_F"
                     /note="color: ?; sequence: 
                     aggggttatgctatgcataccaagcttACCGGTAGTGTTAGACCTG"
     primer_bind     10702..10730
                     /label=kn57
                     /note="pRS305_T1119C_R"
                     /note="color: ?; sequence: ccatcaagaacgaGttccctacattgaag"
     primer_bind     complement(10807..10829)
                     /label=kn1330
                     /note="Leu2_600_R"
                     /note="color: ?; sequence: gatatcaccaaacatgttgctgg"
     primer_bind     11267..11288
                     /label=kn1363
                     /note="Leu2T_F"
                     /note="color: ?; sequence: GGGTAGACGAAACTATATACGC"
     primer_bind     11423..11443
                     /label=kn2442
                     /note="leu2t_end_F"
                     /note="color: ?; sequence: caccacacaaaaagttaggtg"
     primer_bind     complement(11465..11488)
                     /label=kn1364
                     /note="Leu2T_R"
                     /note="color: ?; sequence: CCTCCAATATCAAATTAGGAATCG"
     primer_bind     complement(11525..11546)
                     /label=kn2435
                     /note="amp-Avr-isce-leu2_R"
                     /note="color: ?; sequence: 
                     cgcctagggataacagggtaaTGGTCAGGTCATTGAgtgtt"
     primer_bind     complement(11525..11544)
                     /label=kn2461
                     /note="NSF1end_F"
                     /note="color: ?; sequence: TGGTCAGGTCATTGAgtgtt"
     primer_bind     11527..11548
                     /label=kn1331
                     /note="Leu2_600skip_F"
                     /note="color: ?; sequence: 
                     cagcaacatgtttggtgatatcactcaatgacctgaccatttg"
     primer_bind     complement(11528..11548)
                     /label=kn2476
                     /note="Leu2T-kpnNhe_R"
                     /note="color: ?; sequence: 
                     taggtaccggaggtgctagcaaatggtcaggtcattgagt"
     primer_bind     complement(11651..11684)
                     /label=kn529*
                     /note="pRS_markerout_Sac2-Hind3_F"
                     /note="color: ?; sequence: 
                     caccgcggaagcttagcagattgtactgagagtgcacc"
     primer_bind     complement(11651..11674)
                     /label=kn504
                     /note="pRS_markerout_F"
                     /note="color: ?; sequence: 
                     cgcatagaaattgcatcaacgcatatagcgctagcAGATTGTACTGAGAGTGCACC"
     primer_bind     complement(11651..11671)
                     /label=kn84
                     /note="JW411nest_F"
                     /note="color: ?; sequence: gagattgtactgagagtgcacc"
     primer_bind     complement(11652..11674)
                     /label=kn104
                     /note="pRS_Mark_ENX_F"
                     /note="color: ?; sequence: 
                     tgaattcgcggccgcttctagaagattgtactgagagtgcac"
     primer_bind     complement(11665..11684)
                     /label=kn1198
                     /note="Ura3pro_F"
                     /note="color: ?; sequence: gcggcatcagagcagattgt"
     primer_bind     complement(11670..11689)
                     /label=kn374
                     /note="tonBt-LacZA_R"
                     /note="color: ?; sequence: 
                     CAATTGGTTCAGACGCTACTTTTTTACTATGCGGCATCAGAGCAG"
     primer_bind     complement(11765..11782)
                     /label=kn2299
                     /note="pEX-K4J1_R"
                     /note="color: ?; sequence: AGACGGTCACAGCTTGTC"
     primer_bind     11767..11783
                     /label=kn1157
                     /note="GalK-start-pLlac_R"
                     /note="color: ?; sequence: 
                     GAGTGGCAGGGTAacgcgtacatccatccattttcctcctctgtgctcagtatcttgtt
                     atccgctcacaatgtcaagctgtgaccgtctc"
     primer_bind     11775..11790
                     /label=hm226
                     /note="pCSUII for pT7SU G.A. btm,26"
                     /note="color: ?; sequence: GCCAATCCGGGACCGTCTCCGGGAGC"
     primer_bind     11793..11813
                     /label=kn696
                     /note="pET-SQ2_R"
                     /note="color: ?; sequence: catgtgtcagaggttttcacc"
     primer_bind     complement(11804..11823)
                     /label=kn1333
                     /note="leu2-dPro-term_R"
                     /note="color: ?; sequence: 
                     ttagaatggtatatccttgagctccaccgcggTGATGACGGTGAAAACC"
     primer_bind     complement(11805..11823)
                     /label=kn1344
                     /note="leu2-TAdPro-term_R"
                     /note="color: ?; sequence: 
                     ttagaatggtatatccttGAAATATATATAgagctccaccgcggTGATGACGGTGAAAA
                     C"
     primer_bind     join(11824..11833,1..8)
                     /label=kn2354
                     /note="pSC101_SapI_F"
                     /note="color: ?; sequence: 
                     gtttgCGGCCagctcttcAAACGCGCGAGACGAAAG"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt ttttcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 ggcagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctcgaaatt
     2761 aaccctcact aaagggaaca aaagctggta ccgggcccga attctctcct tctcttaggt
     2821 ggcagagcag gtggagggtc gaccatacta gtttcaaaaa ttcttacttt ttttttggat
     2881 ggacgcaaag aagtttaata atcatattac atggcattac caccatatac atatccatat
     2941 acatatccat atctaatctt acttatatgt tgtggaaatg taaagagccc cattatctta
     3001 gcctaaaaaa accttctctt tggaactttc agtaatacgc ttaactgctc attgctatat
     3061 tgaagtacgg attagaagcc gccgagcggg tgacagccct ccgaaggaag actctcctcc
     3121 gtgcgtcctc gtcttcaccg gtcgcgttcc tgaaacgcag atgtgcctcg cgccgcactg
     3181 ctccgaacaa taaagattct acaatactag cttttatggt tatgaagagg aaaaattggc
     3241 agtaacctgg ccccacaaac cttcaaatga acgaatcaaa ttaacaacca taggatgata
     3301 atgcgattag ttttttagcc ttatttctgg ggtaattaat cagcgaagcg atgatttttg
     3361 atctattaac agatatataa atgcaaaaac tgcataacca ctttaactaa tactttcaac
     3421 attttcggtt tgtattactt cttattcaaa tgtaataaaa gtatcaacaa aaaattgtta
     3481 atatacctct atactttaac gtcaaggaga aaaaaccagc gctcttcgat gtcgcataga
     3541 tgctacgttc tctttgaatt aaaacgacgg ggtgaacgta gagcgtgttt ttggggctat
     3601 gctgtgaata aaccacagag cgggacagaa cgtggcattc acgccgaaat ctttagcatt
     3661 agaaaagtcg aagaatacct gcgcgacaac cccggacaat tcacgataaa ttggtactca
     3721 tcctggagtc cttgtgcaga ttgcgctgaa aagatcttag aatggtataa ccaggagctg
     3781 cgggggaacg gccacacttt gaaaatcgaa gcttgcaaac tctattacga gaaaaatgcg
     3841 aggaatcaaa ttgggctgca aaacctcaga gataacgggg ttgggttgaa tgtaggcgga
     3901 ggtggcagtg gcgggggtgg ctcgggtggg ggaggcagtg gcggaggtgg cagtggcggg
     3961 ggtggctcgg gtgggggagg atccatggac aagaagtact ccattgggct cgctatcggc
     4021 acaaacagcg tcggttgggc cgtcattacg gacgagtaca aggtgccgag caaaaaattc
     4081 aaagttctgg gcaataccga tcgccacagc ataaagaaga acctcattgg cgccctcctg
     4141 ttcgactccg gggagacggc cgaagccacg cggctcaaaa gaacagcacg gcgcagatat
     4201 acccgcagaa agaatcggat ctgctacctg caggagatct ttagtaatga gatggctaag
     4261 gtggatgact ctttcttcca taggctggag gagtcctttt tggtggagga ggataaaaag
     4321 cacgagcgcc acccaatctt tggcaatatc gtggacgagg tggcgtacca tgaaaagtac
     4381 ccaaccatat atcatctgag gaagaagctt gtagacagta ctgataaggc tgacttgcgg
     4441 ttgatctatc tcgcgctggc gcatatgatc aaatttcggg gacacttcct catcgagggg
     4501 gacctgaacc cagacaacag cgatgtcgac aaactcttta tccaactggt tcagacttac
     4561 aatcagcttt tcgaagagaa cccgatcaac gcatccggag ttgacgccaa agcaatcctg
     4621 agcgctaggc tgtccaaatc ccggcggctc gaaaacctca tcgcacagct ccctggggag
     4681 aagaagaacg gcctgtttgg taatcttatc gccctgtcac tcgggctgac ccccaacttt
     4741 aaatctaact tcgacctggc cgaagatgcc aagcttcaac tgagcaaaga cacctacgat
     4801 gatgatctcg acaatctgct ggcccagatc ggcgaccagt acgcagacct ttttttggcg
     4861 gcaaagaacc tgtcagacgc cattctgctg agtgatattc tgcgagtgaa cacggagatc
     4921 accaaagctc cgctgagcgc tagtatgatc aagcgctatg atgagcacca ccaagacttg
     4981 actttgctga aggcccttgt cagacagcaa ctgcctgaga agtacaagga aattttcttc
     5041 gatcagtcta aaaatggcta cgccggatac attgacggcg gagcaagcca ggaggaattt
     5101 tacaaattta ttaagcccat cttggaaaaa atggacggca ccgaggagct gctggtaaag
     5161 cttaacagag aagatctgtt gcgcaaacag cgcactttcg acaatggaag catcccccac
     5221 cagattcacc tgggcgaact gcacgctatc ctcaggcggc aagaggattt ctaccccttt
     5281 ttgaaagata acagggaaaa gattgagaaa atcctcacat ttcggatacc ctactatgta
     5341 ggccccctcg cccggggaaa ttccagattc gcgtggatga ctcgcaaatc agaagagacc
     5401 atcactccct ggaacttcga ggaagtcgtg gataaggggg cctctgccca gtccttcatc
     5461 gaaaggatga ctaactttga taaaaatctg cctaacgaaa aggtgcttcc taaacactct
     5521 ctgctgtacg agtacttcac agtttataac gagctcacca aggtcaaata cgtcacagaa
     5581 gggatgagaa agccagcatt cctgtctgga gagcagaaga aagctatcgt ggacctcctc
     5641 ttcaagacga accggaaagt taccgtgaaa cagctcaaag aagactattt caaaaagatt
     5701 gaatgtttcg actctgttga aatcagcgga gtggaggatc gcttcaacgc atccctggga
     5761 acgtatcacg atctcctgaa aatcattaaa gacaaggact tcctggacaa tgaggagaac
     5821 gaggacattc ttgaggacat tgtcctcacc cttacgttgt ttgaagatag ggagatgatt
     5881 gaagaacgct tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa acagctcaag
     5941 aggcgccgat atacaggatg ggggcggctg tcaagaaaac tgatcaatgg gatccgagac
     6001 aagcagagtg gaaagacaat cctggatttt cttaagtccg atggatttgc caaccggaac
     6061 ttcatgcagt tgatccatga tgactctctc acctttaagg aggacatcca gaaagcacaa
     6121 gtttctggcc agggggacag tcttcacgag cacatcgcta atcttgcagg tagcccagct
     6181 atcaaaaagg gaatactgca gaccgttaag gtcgtggatg aactcgtcaa agtaatggga
     6241 aggcataagc ccgagaatat cgttatcgag atggcccgag agaaccaaac tacccagaag
     6301 ggacagaaga acagtaggga aaggatgaag aggattgaag agggtataaa agaactgggg
     6361 tcccaaatcc ttaaggaaca cccagttgaa aacacccagc ttcagaatga gaagctctac
     6421 ctgtactacc tgcagaacgg cagggacatg tacgtggatc aggaactgga catcaatcgg
     6481 ctctccgact acgacgtgga tcatatcgtg ccccagtctt ttctcaaaga tgattctatt
     6541 gataataaag tgttgacaag atccgataaa aatagaggga agagtgataa cgtcccctca
     6601 gaagaagttg tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc caaactgatc
     6661 acacaacgga agttcgataa tctgactaag gctgaacgag gtggcctgtc tgagttggat
     6721 aaagccggct tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa gcacgtggcc
     6781 caaattctcg attcacgcat gaacaccaag tacgatgaaa atgacaaact gattcgagag
     6841 gtgaaagtta ttactctgaa gtctaagctg gtctcagatt tcagaaagga ctttcagttt
     6901 tataaggtga gagagatcaa caattaccac catgcgcatg atgcctacct gaatgcagtg
     6961 gtaggcactg cacttatcaa aaaatatccc aagcttgaat ctgaatttgt ttacggagac
     7021 tataaagtgt acgatgttag gaaaatgatc gcaaagtctg agcaggaaat aggcaaggcc
     7081 accgctaagt acttctttta cagcaatatt atgaattttt tcaagaccga gattacactg
     7141 gccaatggag agattcggaa gcgaccactt atcgaaacaa acggagaaac aggagaaatc
     7201 gtgtgggaca agggtaggga tttcgcgaca gtccggaagg tcctgtccat gccgcaggtg
     7261 aacatcgtta aaaagaccga agtacagacc ggaggcttct ccaaggaaag tatcctcccg
     7321 aaaaggaaca gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa gaaatacggc
     7381 ggattcgatt ctcctacagt cgcttacagt gtactggttg tggccaaagt ggagaaaggg
     7441 aagtctaaaa aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat ggagcgatca
     7501 agcttcgaaa aaaaccccat cgactttctc gaggcgaaag gatataaaga ggtcaaaaaa
     7561 gacctcatca ttaagcttcc caagtactct ctctttgagc ttgaaaacgg ccggaaacga
     7621 atgctcgcta gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc ctctaaatac
     7681 gttaatttct tgtatctggc cagccactat gaaaagctca aagggtctcc cgaagataat
     7741 gagcagaagc agctgttcgt ggaacaacac aaacactacc ttgatgagat catcgagcaa
     7801 ataagcgaat tctccaaaag agtgatcctc gccgacgcta acctcgataa ggtgctttct
     7861 gcttacaata agcacaggga taagcccatc agggagcagg cagaaaacat tatccacttg
     7921 tttactctga ccaacttggg cgcgcctgca gccttcaagt acttcgacac caccatagac
     7981 agaaagcggt acacctctac aaaggaggtc ctggacgcca cactgattca tcagtcaatt
     8041 acggggctct atgaaacaag aatcgacctc tctcagctcg gtggagacag cagggctgac
     8101 cccaagaaga agaggaaggt gggtaccatg accaaccttt ccgacatcat agagaaggaa
     8161 acaggcaaac agttggtcat ccaagagtcg atactcatgc ttcctgaaga agttgaggag
     8221 gtcattggga ataagccgga aagtgacatt ctcgtacaca ctgcgtatga tgagagcacc
     8281 gatgagaacg tgatgctgct cacgtcagat gccccagagt acaaaccctg ggctctggtg
     8341 attcaggact ctaatggaga gaacaagatc aagatgctat agtctagagg ctccggataa
     8401 ggatcctaat aagcgaattt cttatgattt atgattttta ttattaaata agttataaaa
     8461 aaaataagtg tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc
     8521 ttgagtaact ctttcctgta ggtcaggttg ctttctcagg tatagcatga ggtcgctctt
     8581 attgaccaca cctctaccgg catgccgagc aaatgcctgc aaatcgctcc cgggcaaaaa
     8641 accccctcaa gacccgttta gaggccccaa ggggttatgc tatgcatacc aagcttccgc
     8701 ggtggagctc caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac
     8761 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     8821 ccttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     8881 gcagcctgaa tggcgaatgg cgcgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     8941 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     9001 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     9061 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     9121 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     9181 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     9241 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     9301 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     9361 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatcgaccct
     9421 cgaggagaac ttctagtata tccacatacc taatattatt gccttattaa aaatggaatc
     9481 ggaacaatta catcaaaatc cacattctct tcaaaatcaa ttgtcctgta cttccttgtt
     9541 catgtgtgtt caaaaacgtt atatttatag gataattata ctctatttct caacaagtaa
     9601 ttggttgttt ggccgagcgg tctaaggcgc ctgattcaag aaatatcttg accgcagtta
     9661 actgtgggaa tactcaggta tcgtaagatg caagagttcg aatctcttag caaccattat
     9721 ttttttcctc aacataacga gaacacacag gggcgctatc gcacagaatc aaattcgatg
     9781 actggaaatt ttttgttaat ttcagaggtc gcctgacgca tatacctttt tcaactgaaa
     9841 aattgggaga aaaaggaaag gtgagaggcc ggaaccggct tttcatatag aatagagaag
     9901 cgttcatgac taaatgcttg catcacaata cttgaagttg acaatattat ttaaggacct
     9961 attgtttttt ccaataggtg gttagcaatc gtcttacttt ctaacttttc ttacctttta
    10021 catttcagca atatatatat atatttcaag gatataccat tctaatgtct gcccctatgt
    10081 ctgcccctaa gaagatcgtc gttttgccag gtgaccacgt tggtcaagaa atcacagccg
    10141 aagccattaa ggttcttaaa gctatttctg atgttcgttc caatgtcaag ttcgatttcg
    10201 aaaatcattt aattggtggt gctgctatcg atgctacagg tgtcccactt ccagatgagg
    10261 cgctggaagc ctccaagaag gttgatgccg ttttgttagg tgctgtgggt ggtcctaaat
    10321 ggggtaccgg tagtgttaga cctgaacaag gtttactaaa aatccgtaaa gaacttcaat
    10381 tgtacgccaa cttaagacca tgtaactttg catccgactc tcttttagac ttatctccaa
    10441 tcaagccaca atttgctaaa ggtactgact tcgttgttgt cagagaatta gtgggaggta
    10501 tttactttgg taagagaaag gaagacgatg gtgatggtgt cgcttgggat agtgaacaat
    10561 acaccgttcc agaagtgcaa agaatcacaa gaatggccgc tttcatggcc ctacaacatg
    10621 agccaccatt gcctatttgg tccttggata aagctaatgt tttggcctct tcaagattat
    10681 ggagaaaaac tgtggaggaa accatcaaga acgaattccc tacattgaag gttcaacatc
    10741 aattgattga ttctgccgcc atgatcctag ttaagaaccc aacccaccta aatggtatta
    10801 taatcaccag caacatgttt ggtgatatca tctccgatga agcctccgtt atcccaggtt
    10861 ccttgggttt gttgccatct gcgtccttgg cctctttgcc agacaagaac accgcatttg
    10921 gtttgtacga accatgccac ggttctgctc cagatttgcc aaagaataag gttgacccta
    10981 tcgccactat cttgtctgct gcaatgatgt tgaaattgtc attgaacttg cctgaagaag
    11041 gtaaggccat tgaagatgca gttaaaaagg ttttggatgc aggtatcaga actggtgatt
    11101 taggtggttc caacagtacc accgaagtcg gtgatgctgt cgccgaagaa gttaagaaaa
    11161 tccttgctta aaaagattct ctttttttat gatatttgta cataaacttt ataaatgaaa
    11221 ttcataatag aaacgacacg aaattacaaa atggaatatg ttcatagggt agacgaaact
    11281 atatacgcaa tctacataca tttatcaaga aggagaaaaa ggaggatagt aaaggaatac
    11341 aggtaagcaa attgatacta atggctcaac gtgataagga aaaagaattg cactttaaca
    11401 ttaatattga caaggaggag ggcaccacac aaaaagttag gtgtaacaga aaatcatgaa
    11461 actacgattc ctaatttgat attggaggat tttctctaaa aaaaaaaaaa tacaacaaat
    11521 aaaaaacact caatgacctg accatttgat ggagtttaag tcaatacctt cttgaaccat
    11581 ttcccataat ggtgaaagtt ccctcaagaa ttttactctg tcagaaacgg ccttacgacg
    11641 tagtcgatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
    11701 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
    11761 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
    11821 ccgaaacgcg cga
//
