LOCUS       pUT_mNF-Cas13d-T       11901 bp DNA     circular SYN 22-JAN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 11901)
  AUTHORS   Yiming Wan
  TITLE     Direct Submission
  JOURNAL   Exported Sep 26, 2024 from SnapGene Viewer 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11901
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     primer_bind     complement(1..25)
                     /label=pRIME-mLin-GFP-1.HiF
     primer_bind     1..25
                     /label=pRIME-mLin-GFP-2.HiR
     primer_bind     complement(6..24)
                     /label=pUT-AmpR-addon-2.HiF
     primer_bind     6..23
                     /label=nvn.1.19
     primer_bind     complement(6..23)
                     /label=nvn.1.18
     rep_origin      15..603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     196..225
                     /label=Ori-Split.Rev
     primer_bind     complement(196..225)
                     /label=Ori-Split.fwd
     primer_bind     424..453
                     /label=HiFiOriSplit.Rev
     primer_bind     complement(424..453)
                     /label=HifiOriSplit.Fwd
     primer_bind     497..516
                     /label=pU3_pciI_seq_R
     primer_bind     516..537
                     /label=pHR-mLin-GFP-Cas13d-3.HiF
     primer_bind     complement(586..605)
                     /label=pPT2-backbone-1_v2.HiR
     primer_bind     complement(590..653)
                     /label=pRIME-mLin-Twist-2.HiF
     primer_bind     complement(590..609)
                     /label=pPT2-PuroR-backbone-1.HiR
     primer_bind     complement(600..623)
                     /label=pPT2-Backbone-1.HiR
     primer_bind     606..625
                     /label=pHR-mLin-mCherry-Cas13d-3.HiF
     primer_bind     608..661
                     /label=mLin-BACH1-SGC.F
     misc_feature    complement(608..643)
                     /label=FRT (minimal)
     primer_bind     625..653
                     /label=pRIME-mLin-Twist-3.HiFv2
     primer_bind     628..643
                     /label=proL-NF-GFPv4-2.2.HiF
     primer_bind     complement(634..654)
                     /label=pPT-eSpCas9-sgSca-3.HiFv2
     primer_bind     634..653
                     /label=pPT-mLin-Backbone-2.HiF
     primer_bind     complement(634..653)
                     /label=pPT-mLin-Backbone-1.HiR
     primer_bind     complement(634..653)
                     /label=pPT-eSpCas9-sgSca-3.HiF
     primer_bind     complement(639..677)
                     /label=CRISPRa-donor-vector.HiR
     primer_bind     641..661
                     /label=pPT-mLin-Split-2.HiR
     primer_bind     complement(641..661)
                     /label=pPT-mLin-Split-1.HiR
     primer_bind     complement(642..679)
                     /label=pPT-mPF-GFP-BACH1-1.HiR
     primer_bind     complement(643..660)
                     /label=FRT3-pUt-DC.R
     primer_bind     complement(644..661)
                     /label=pUT-mPF-GFPv2-1.HiR
     primer_bind     644..661
                     /label=pUT-mPF-GFPv2-2.HiF
     primer_bind     complement(644..659)
                     /label=CRISPRa-donor-vector.HiR
     promoter        661
                     /label=CMV-D2i Promoter
     primer_bind     complement(662..679)
                     /label=pUT-mPF-GFPv2-1.HiR
     primer_bind     662..679
                     /label=pUT-mPF-GFPv2-2.HiF
     primer_bind     complement(662..678)
                     /label=FRT3-pUt-DC.R
     primer_bind     complement(680..697)
                     /label=pUT-mPF-GFPv2-1.HiR
     primer_bind     680..697
                     /label=mLin-BACH1-SGC.F
     primer_bind     680..697
                     /label=pUT-mPF-GFPv2-2.HiF
     primer_bind     complement(680..696)
                     /label=FRT3-pUt-DC.R
     primer_bind     complement(680..695)
                     /label=CRISPRa-donor-vector.HiR
     primer_bind     680..689
                     /label=pPT-mLin-Backbone-2.HiF
     primer_bind     680..689
                     /label=pRIME-mLin-Twist-3.HiFv2
     primer_bind     complement(698..715)
                     /label=pUT-mPF-GFPv2-1.HiR
     primer_bind     698..715
                     /label=mLin-BACH1-SGC.F
     primer_bind     698..715
                     /label=pUT-mPF-GFPv2-2.HiF
     primer_bind     complement(698..714)
                     /label=FRT3-pUt-DC.R
     primer_bind     complement(698..713)
                     /label=CRISPRa-donor-vector.HiR
     primer_bind     698..707
                     /label=pPT-mLin-Backbone-2.HiF
     primer_bind     698..707
                     /label=pRIME-mLin-Twist-3.HiFv2
     primer_bind     complement(716..733)
                     /label=pUT-mPF-GFPv2-1.HiR
     primer_bind     716..733
                     /label=mLin-BACH1-SGC.F
     primer_bind     716..733
                     /label=pUT-mPF-GFPv2-2.HiF
     primer_bind     complement(716..732)
                     /label=FRT3-pUt-DC.R
     primer_bind     complement(716..731)
                     /label=CRISPRa-donor-vector.HiR
     primer_bind     716..725
                     /label=pPT-mLin-Backbone-2.HiF
     primer_bind     716..725
                     /label=pRIME-mLin-Twist-3.HiFv2
     primer_bind     complement(734..751)
                     /label=pUT-mPF-GFPv2-1.HiR
     primer_bind     734..751
                     /label=mLin-BACH1-SGC.F
     primer_bind     734..751
                     /label=pUT-mPF-GFPv2-2.HiF
     primer_bind     complement(734..750)
                     /label=FRT3-pUt-DC.R
     primer_bind     complement(734..749)
                     /label=CRISPRa-donor-vector.HiR
     primer_bind     734..743
                     /label=pPT-mLin-Backbone-2.HiF
     primer_bind     734..743
                     /label=pRIME-mLin-Twist-3.HiFv2
     primer_bind     complement(752..769)
                     /label=pUT-mPF-GFPv2-1.HiR
     primer_bind     752..769
                     /label=mLin-BACH1-SGC.F
     primer_bind     752..769
                     /label=pUT-mPF-GFPv2-2.HiF
     primer_bind     complement(752..768)
                     /label=FRT3-pUt-DC.R
     primer_bind     complement(752..767)
                     /label=CRISPRa-donor-vector.HiR
     primer_bind     752..761
                     /label=pPT-mLin-Backbone-2.HiF
     primer_bind     752..761
                     /label=pRIME-mLin-Twist-3.HiFv2
     primer_bind     complement(770..787)
                     /label=pUT-mPF-GFPv2-1.HiR
     primer_bind     770..787
                     /label=mLin-BACH1-SGC.F
     primer_bind     770..787
                     /label=pUT-mPF-GFPv2-2.HiF
     primer_bind     complement(770..786)
                     /label=FRT3-pUt-DC.R
     primer_bind     complement(770..785)
                     /label=CRISPRa-donor-vector.HiR
     primer_bind     770..779
                     /label=pPT-mLin-Backbone-2.HiF
     primer_bind     770..779
                     /label=pRIME-mLin-Twist-3.HiFv2
     primer_bind     complement(788..805)
                     /label=pUT-mPF-GFPv2-1.HiR
     primer_bind     788..805
                     /label=mLin-BACH1-SGC.F
     primer_bind     788..805
                     /label=pUT-mPF-GFPv2-2.HiF
     primer_bind     complement(788..804)
                     /label=FRT3-pUt-DC.R
     primer_bind     complement(788..803)
                     /label=CRISPRa-donor-vector.HiR
     primer_bind     788..797
                     /label=pPT-mLin-Backbone-2.HiF
     primer_bind     788..797
                     /label=pRIME-mLin-Twist-3.HiFv2
     primer_bind     complement(806..823)
                     /label=pUT-mPF-GFPv2-1.HiR
     primer_bind     806..823
                     /label=mLin-BACH1-SGC.F
     primer_bind     806..823
                     /label=pUT-mPF-GFPv2-2.HiF
     primer_bind     complement(806..822)
                     /label=FRT3-pUt-DC.R
     primer_bind     complement(806..821)
                     /label=CRISPRa-donor-vector.HiR
     primer_bind     806..815
                     /label=pPT-mLin-Backbone-2.HiF
     primer_bind     806..815
                     /label=pRIME-mLin-Twist-3.HiFv2
     primer_bind     complement(824..841)
                     /label=pUT-mPF-GFPv2-1.HiR
     primer_bind     824..841
                     /label=mLin-BACH1-SGC.F
     primer_bind     824..841
                     /label=pUT-mPF-GFPv2-2.HiF
     primer_bind     complement(824..840)
                     /label=FRT3-pUt-DC.R
     primer_bind     complement(824..839)
                     /label=CRISPRa-donor-vector.HiR
     primer_bind     824..833
                     /label=pPT-mLin-Backbone-2.HiF
     primer_bind     824..833
                     /label=pRIME-mLin-Twist-3.HiFv2
     primer_bind     complement(842..878)
                     /label=pPT-mPF-GFP-BACH1-1.HiR
     primer_bind     complement(842..859)
                     /label=pUT-mPF-GFPv2-1.HiR
     primer_bind     842..859
                     /label=mLin-BACH1-SGC.F
     primer_bind     842..859
                     /label=pUT-mPF-GFPv2-2.HiF
     primer_bind     complement(842..858)
                     /label=FRT3-pUt-DC.R
     primer_bind     complement(842..857)
                     /label=CRISPRa-donor-vector.HiR
     primer_bind     842..851
                     /label=pPT-mLin-Backbone-2.HiF
     primer_bind     842..851
                     /label=pRIME-mLin-Twist-3.HiFv2
     primer_bind     860..885
                     /label=CMV-SgrDI-mut-f
     primer_bind     complement(860..879)
                     /label=pUT-mPF-GFPv2-1.HiR
     primer_bind     complement(860..879)
                     /label=KRASB-HSV.R
     primer_bind     860..878
                     /label=pUT-mPF-GFPv2-2.HiF
     primer_bind     860..877
                     /label=mLin-BACH1-SGC.F
     primer_bind     complement(860..876)
                     /label=FRT3-pUt-DC.R
     primer_bind     complement(860..875)
                     /label=CRISPRa-donor-vector.HiR
     primer_bind     860..869
                     /label=pPT-mLin-Backbone-2.HiF
     primer_bind     860..869
                     /label=pRIME-mLin-Twist-3.HiFv2
     primer_bind     864..883
                     /label=nvn.1.16
     primer_bind     872..939
                     /label=MluI-CMV
     primer_bind     875..899
                     /label=proL-NF-GFPv4-2.HiF
     primer_bind     875..899
                     /label=proL-NF-GFPv4-2.2.HiF
     primer_bind     complement(875..894)
                     /label=pRIME-mLin-GFP-1.HiR
     primer_bind     complement(875..894)
                     /label=GFP_Linearizer_FRT.HiR
     primer_bind     875..894
                     /label=pRIME-mLin-GFP-2.HiF
     primer_bind     877..915
                     /label=CMV-D2i-SgrDI-f
     primer_bind     877..903
                     /label=NsiI-D2ir-f
     promoter        880..1549
                     /label=CMV-D2i Promoter
     enhancer        880..1259
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     880..920
                     /label=SalI-CMV-D2ir-f
     primer_bind     880..905
                     /label=CMV-enhancer-upstream-seq
     primer_bind     1183..1202
                     /label=Put7.0_Intermediate_RFP_HiF
     primer_bind     1204..1226
                     /label=protrap_Fluo-AAVS1_RFP.HiFa
     primer_bind     1207..1226
                     /label=protrap_Fluo-AAVS1_RFP.HiF
     primer_bind     1207..1226
                     /label=pUt7.0M-fluo-red.HiR
     primer_bind     complement(1216..1244)
                     /label=pAAVS1-Rev(2)
     primer_bind     complement(1216..1244)
                     /label=pAAVS1_rev(3)
     primer_bind     1225..1277
                     /label=FRT-CMV_Promoter-f
     primer_bind     complement(1225..1259)
                     /label=HR(AAVS1)-RFP.HiR
     primer_bind     1260..1280
                     /label=CMV_Promoter_fwd
                     /note="3'Tm=56.1 3'Ta(annealing temp)=55.0"
     primer_bind     1274..1293
                     /label=CMV-seq.fwd
                     /note="Sequencing primer from CMV"
     primer_bind     complement(1439..1458)
                     /label=CMV_Promoter_rev
                     /note="3'Tm=55.0 3'Ta(annealing temp)=55.0"
     TATA_signal     1449..1455
     protein_bind    1459..1477
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     TATA_signal     1478..1484
                     /label=Initiator Element
     protein_bind    1485..1503
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(1492..1515)
                     /label=NF-BACH1 head internal v1
     primer_bind     1509..1526
                     /label=pCMV-TetR-mCherry-Cas13d-2.HiF
     primer_bind     1510..1526
                     /label=pPT-mLin-mCherry-Split-Cas13d-2.HiF
     primer_bind     complement(1537..1556)
                     /label=NF-head-fwd-v2
     primer_bind     1549..1573
                     /label=bG-intron-seq-fwd
     primer_bind     complement(1557..1581)
                     /label=NF-head-fwd
     primer_bind     complement(1617..1636)
                     /label=RIME-2-1.HiFv2
     primer_bind     complement(1625..1640)
                     /label=PPT-to-PUT change-2.HiR
     primer_bind     1625..1640
                     /label=PPT-to-PUT change-1.HiF
     primer_bind     complement(1625..1640)
                     /label=PPT-to-PUT change-2.HiR
     primer_bind     complement(1625..1640)
                     /label=PPT-to-PUT change-2.HiR
     primer_bind     complement(1625..1640)
                     /label=RIME-1.1
     primer_bind     complement(1625..1640)
                     /label=PPT-to-PUT change-2.HiR
     primer_bind     complement(1625..1640)
                     /label=PPT-to-PUT change-2.HiR (2)
     primer_bind     complement(1625..1640)
                     /label=PPT-to-PUT change-2.HiR (2) (2)
     primer_bind     complement(1625..1640)
                     /label=PPT-to-PUT change-2.HiR (3)
     primer_bind     complement(1625..1640)
                     /label=pPT2-Cas13-EGFP-KD-test-1.HiR
     primer_bind     complement(1625..1640)
                     /label=RIME-1.1-2
     intron          1686..2258
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     1686..1705
                     /label=pCMV-TetR-mCherry-Cas13d-2.HiF
     primer_bind     complement(2216..2235)
                     /label=p23_kzkmut_pREV
     primer_bind     2234..2258
                     /label=mLin-D2t-Cas13d-1.HiF
     primer_bind     complement(2234..2258)
                     /label=bG-intron-seq-rev
     primer_bind     complement(2249..2278)
                     /label=etRFP-D2ir-bb.R
     primer_bind     2252..2274
                     /label=Intron-SbfI-f
     primer_bind     2267..2292
                     /label=hTetR-KasI-f
     primer_bind     2267..2284
                     /label=KasI-TNP2aG-f
     primer_bind     2269..2301
                     /label=KpnI-KasI-KozIN-rtTA-OL-f
     primer_bind     2269..2292
                     /label=mutKpnI-Koz-rtTA-2ndOL-f
     primer_bind     2270..2301
                     /label=KozIN-rtTA-1stOL-f
     CDS             2276..2899
                     /codon_start=1
                     /label=hTetR
                     /translation="MGSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGG"
     primer_bind     2276..2301
                     /label=hTetR-seq-fwd
     primer_bind     complement(2288..2311)
                     /label=hTetR-N-Term-SEQ-r
     primer_bind     complement(2336..2357)
                     /label=NF-BACH1 head internal v2
     primer_bind     2492..2511
                     /label=nvn.1.12
     primer_bind     complement(2492..2511)
                     /label=nvn.1.13
     primer_bind     2606..2625
                     /label=nvn.1.12-2
     primer_bind     complement(2606..2625)
                     /label=nvn.1.13-2
     primer_bind     2866..2887
                     /label=hTetR-C-Term-Seq-f
     primer_bind     complement(2880..2929)
                     /label=NLS-P2a-1stOL-r
     primer_bind     2880..2899
                     /label=pPT-mLin-target.HiF
     primer_bind     complement(2880..2899)
                     /label=pPT-mLin-scaffold.HiR
     primer_bind     complement(2899..2924)
                     /label=hTetR-seq-rev
     CDS             2900..2917
                     /label=NLS
     primer_bind     2916..2941
                     /label=TG1.31.14
     primer_bind     complement(2918..2941)
                     /label=TG1.16.4
     primer_bind     2918..2941
                     /label=TG1.31.2
     primer_bind     2918..2941
                     /label=TG1.31.10
     primer_bind     2918..2939
                     /label=TG1.13.5
     CDS             2927..2983
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(2963..2986)
                     /label=ATG-P2a.R
     primer_bind     complement(2963..2986)
                     /label=P2a-EtagRFP.Ra
     primer_bind     2963..2983
                     /label=P2a-pUt-DC.F
     primer_bind     complement(2963..2983)
                     /label=TG1.7.6
     primer_bind     complement(2964..2993)
                     /label=pPT-mLin-scaffold-GFP-GB.HiR
     primer_bind     complement(2964..2989)
                     /label=pUT-mLin-Twsitv2-2.HiF
     primer_bind     complement(2964..2986)
                     /label=pUt-NF-GFP_NVN_no-Bach1_HiFi-Ra
     primer_bind     complement(2964..2986)
                     /label=CRISPRa-donor-vector.HiR-2
     primer_bind     complement(2964..2986)
                     /label=put_NF_BACH1_P2A_GFP-1.HiR
     primer_bind     complement(2964..2986)
                     /label=TG1.31.1
     primer_bind     complement(2964..2986)
                     /label=TG1.31.9
     primer_bind     complement(2964..2983)
                     /label=P2a-OL-r
     primer_bind     complement(2964..2983)
                     /label=mLin-Cas13d-1.HiF
     primer_bind     complement(2964..2983)
                     /label=pPT-mLin-Backbone-2.HiR
     primer_bind     complement(2964..2983)
                     /label=pPT-D2ir-BACH1_BlastR-1.HiF
     primer_bind     complement(2966..2986)
                     /label=pUt-NF-BACH1.R
     primer_bind     complement(2966..2986)
                     /label=P2a-EtagRFP.R
     primer_bind     complement(2966..2983)
                     /label=pUt-NF.R
     primer_bind     complement(2966..2983)
                     /label=mNF-bb.R
     primer_bind     complement(2969..2983)
                     /label=pUt-NF_NVN_HiFi-Rev
     primer_bind     complement(2969..2983)
                     /label=pUt-NF-GFP_NVN_no-Bach1_HiFi-R
     primer_bind     complement(2971..2996)
                     /label=P2a-NeoGFP.R
     primer_bind     2972..3004
                     /label=2A-NeonGreen.F
     primer_bind     complement(2972..2982)
                     /label=pUT-mLin-Twist-1.HiF
     primer_bind     complement(2973..2982)
                     /label=mLin-denAsCas12a-backbone.HiF
     primer_bind     2974..2993
                     /label=pPT-mLin-target-GFP-GB.HiF
     misc_feature    2984..3691
                     /locus_tag="mCherry"
                     /label=mCherry
                     /note="mCherry"
     primer_bind     complement(2984..3003)
                     /label=mLin-mCherry-Cas13d-1.HiF
     primer_bind     2984..3003
                     /label=mLin-mCherry-Cas13d-2.HiF
     primer_bind     complement(2984..3002)
                     /label=hEGFP-N-Term-Seq-r
     primer_bind     2984..3001
                     /label=HiFi-KRAS-GFP.fwd
     primer_bind     2984..3001
                     /label=pUt-NF-GFP_NVN_no-Bach1_HiFi-F-ZeoRLinker
     primer_bind     2984..2998
                     /label=pUt-NF-GFP_NVN_no-Bach1_HiFi-F
     primer_bind     2985..3004
                     /label=TG1.10.5
     primer_bind     2985..3004
                     /label=TG1.10.5a
     primer_bind     complement(3670..3756)
                     /label=T2a-hEGFP-OL-r
     primer_bind     complement(3670..3703)
                     /label=mLin-Vec.R
     primer_bind     complement(3670..3691)
                     /label=noBach1_FragB-rev_Stop_V1
     primer_bind     complement(3670..3691)
                     /label=pUt-NF-GFP_NVN_Mutagenesis-R
     primer_bind     complement(3671..3691)
                     /label=hEGFP-AgeI-BGH-pA-1stOL-r
     primer_bind     complement(3671..3690)
                     /label=EGFP Fwd SL
     primer_bind     complement(3672..3703)
                     /label=G-Linker-OL-r
     primer_bind     3672..3691
                     
     primer_bind     3672..3691
                     /label=nvn1.1.1
     primer_bind     3672..3691
                     /label=pUT-mLin-mCherry.HiF
     primer_bind     3672..3691
                     /label=pHR-mLin-mCherry-Cas13d-4.HiF
     primer_bind     complement(3672..3691)
                     /label=nvn1.1.4
     primer_bind     complement(3672..3691)
                     /label=TG1.10.6
     primer_bind     complement(3672..3691)
                     /label=pHR-mLin-mCherry-Cas13d-3.HiR
     primer_bind     complement(3672..3691)
                     /label=pUT-mLin-mCherry.HiR
     primer_bind     complement(3680..3691)
                     /label=BamHI-hEGFP-r
     primer_bind     complement(3681..3700)
                     /label=pUT-mLin-Twsit-1.HiF
     primer_bind     3690..3715
                     /label=TG1.31.14
     primer_bind     3692..3715
                     /label=TG1.31.10
     primer_bind     complement(3695..3715)
                     /label=TG1.16.4
     primer_bind     3695..3715
                     /label=TG1.31.2
     primer_bind     3695..3713
                     /label=TG1.13.5
     misc_feature    3701..3757
                     /locus_tag="P2A"
                     /label=P2A
                     /note="P2A"
     primer_bind     3737..3756
                     /label=P2a-pUt-DC.F
     primer_bind     complement(3737..3756)
                     /label=ATG-P2a.R
     primer_bind     complement(3737..3756)
                     /label=P2a-EtagRFP.Ra
     primer_bind     complement(3737..3756)
                     /label=TG1.7.6
     primer_bind     complement(3738..3756)
                     /label=P2a-OL-r
     primer_bind     complement(3738..3756)
                     /label=pUt-NF-GFP_NVN_no-Bach1_HiFi-Ra
     primer_bind     complement(3738..3756)
                     /label=CRISPRa-donor-vector.HiR-2
     primer_bind     complement(3738..3756)
                     /label=put_NF_BACH1_P2A_GFP-1.HiR
     primer_bind     complement(3738..3756)
                     /label=mLin-Cas13d-1.HiF
     primer_bind     complement(3738..3756)
                     /label=TG1.31.1
     primer_bind     complement(3738..3756)
                     /label=pPT-mLin-Backbone-2.HiR
     primer_bind     complement(3738..3756)
                     /label=pUT-mLin-Twsitv2-2.HiF
     primer_bind     complement(3738..3756)
                     /label=TG1.31.9
     primer_bind     complement(3738..3756)
                     /label=pPT-mLin-scaffold-GFP-GB.HiR
     primer_bind     complement(3740..3756)
                     /label=pUt-NF.R
     primer_bind     complement(3740..3756)
                     /label=pUt-NF-BACH1.R
     primer_bind     complement(3740..3756)
                     /label=mNF-bb.R
     primer_bind     complement(3740..3756)
                     /label=P2a-EtagRFP.R
     primer_bind     complement(3743..3756)
                     /label=pUt-NF_NVN_HiFi-Rev
     primer_bind     complement(3743..3756)
                     /label=pUt-NF-GFP_NVN_no-Bach1_HiFi-R
     primer_bind     complement(3745..3756)
                     /label=P2a-NeoGFP.R
     primer_bind     complement(3746..3765)
                     /label=pUT-mLin-Twist-1.HiF
     primer_bind     complement(3747..3764)
                     /label=mLin-denAsCas12a-backbone.HiF
     misc_feature    3755..3760
                     /locus_tag="XmaI"
                     /label=XmaI
     misc_feature    3761..6661
                     /locus_tag="Rfx Cas13d"
                     /label=Rfx Cas13d
     primer_bind     3761..3785
                     /label=pPT-mLin-mCherry-Split-Cas13d-2.HiF
     primer_bind     3761..3780
                     /label=Cas13d-head-seq.F
     misc_feature    4475..4492
                     /locus_tag="HEPN1"
                     /label=HEPN1
     primer_bind     complement(4967..4986)
                     /label=Cas13d-head-seq.R
     primer_bind     4989..5008
                     /label=Cas13d-body-seq.F
     primer_bind     complement(6170..6189)
                     /label=Cas13d-body-seq.R
     primer_bind     6191..6210
                     /label=Cas13d-tail-seq.F
     misc_feature    6332..6349
                     /locus_tag="HEPN2"
                     /label=HEPN2
     primer_bind     complement(6419..6438)
                     /label=mLin-D2t-Cas13d-1.HiR
     primer_bind     6419..6438
                     /label=mLin-D2t-Cas13d-2.HiF
     primer_bind     complement(6640..6661)
                     /label=Cas13d-3UTR-CrEGFP_NoNLS-1.HiF
     primer_bind     complement(6640..6661)
                     /label=Cas13d-3UTR-CrEGFP_NoNLS-1.HiFv2
     primer_bind     complement(6641..6660)
                     /label=Cas13d-tail-seq.R
     misc_feature    6662
                     /locus_tag="SpeI(1)"
                     /label=SpeI(1)
     misc_feature    6663..6667
                     /locus_tag="SpeI"
                     /label=SpeI
     misc_feature    6668..6670
                     /locus_tag="linker from baohui(2)"
                     /label=linker from baohui(2)
     misc_feature    6671
                     /locus_tag="NLS"
                     /label=NLS
     misc_feature    6671
                     /locus_tag="linker from baohui(1)"
                     /label=linker from baohui(1)
     misc_feature    6672..6678
                     /locus_tag="NLS(1)"
                     /label=NLS(1)
     misc_feature    6672..6678
                     /locus_tag="linker from baohui(3)"
                     /label=linker from baohui(3)
     misc_feature    6679..6685
                     /locus_tag="sv40 NLS"
                     /label=sv40 NLS
     misc_feature    6679..6685
                     /locus_tag="linker from baohui"
                     /label=linker from baohui
     misc_feature    6686..6694
                     /locus_tag="linker from baohui(4)"
                     /label=linker from baohui(4)
     misc_feature    6686..6691
                     /locus_tag="NLS(2)"
                     /label=NLS(2)
     misc_feature    6695..6700
                     /locus_tag="linker from baohui(5)"
                     /label=linker from baohui(5)
     misc_feature    6701..6727
                     /locus_tag="c-Myc NLS"
                     /label=c-Myc NLS
     misc_feature    6728..6733
                     /locus_tag="BamHI"
                     /label=BamHI
     misc_feature    6734..6746
                     /locus_tag="3x Flag Tag(3)"
                     /label=3x Flag Tag(3)
     misc_feature    6747..6765
                     /locus_tag="3x Flag Tag(2)"
                     /label=3x Flag Tag(2)
     misc_feature    6766..6790
                     /locus_tag="3x Flag Tag(1)"
                     /label=3x Flag Tag(1)
     primer_bind     complement(6789..6831)
                     /label=13-Triplex-CrRNA-1.HiR
     misc_feature    6791..6799
                     /locus_tag="3x Flag Tag"
                     /label=3x Flag Tag
     misc_feature    6803..6808
                     /locus_tag="BsiWI"
                     /label=BsiWI
     primer_bind     6809..6831
                     /label=Triplex-sgRNA-pA.HiF
     primer_bind     6809..6831
                     /label=mLin-mCherry-miR-EGFP-1.HiF
     primer_bind     6812..6836
                     /label=TetOn-Cas12a-Triplex-1.HiF
     misc_feature    6818..6927
                     /label=Triplex stablier
     primer_bind     6938..6977
                     /label=13-Triplex-CrRNA-1.HiF
     primer_bind     complement(6938..6957)
                     /label=13-Triplex-CrRNA-2.HiR
     primer_bind     complement(6938..6957)
                     /label=mLin-mCherry-miR-EGFP-1.HiR
     primer_bind     6957..6977
                     /label=CrRNA_array_amplify-fwd
     primer_bind     6957..6971
                     /label=pUT-mLin-Twsit-1.HiRa
     primer_bind     6957..6968
                     /label=mLin-denAsCas12a-backbone-2.HiR
     primer_bind     6957..6968
                     /label=pUT-mLin-Twsit-1.HiR
     primer_bind     6957..6968
                     /label=pUT-mLin-mCherry.HiF
     misc_feature    6957
                     /locus_tag="BsiWI"
                     /label=BsiWI
     primer_bind     6959..6978
                     /label=mLin-denAsCas12a-backbone-1.HiR
     primer_bind     7003..7013
                     /label=pUt-NF-GFP_Mutagenesis-F_V2
     primer_bind     7003..7012
                     /label=mLin-Vec.F
     primer_bind     7004..7023
                     
     primer_bind     7004..7023
                     /label=nvn1.1.1
     primer_bind     complement(7004..7023)
                     /label=noBach1_FragB-rev_Stop_V1
     primer_bind     complement(7004..7023)
                     /label=nvn1.1.4
     primer_bind     7009..7040
                     /label=RKIP-TAA-WPRE.F
     primer_bind     7016..7067
                     /label=pUt-NF.F
     misc_feature    7025..7383
                     /label=WPRE
     primer_bind     7025..7048
                     /label=etRFP-D2ir-bb.F
     primer_bind     complement(7066..7085)
                     /label=pPT-mLin-target_v3-1.HiR
     primer_bind     7384..7404
                     /label=NotI-WPRE-DEL-f
     primer_bind     7435..7456
                     /label=bGH-Blst.F
     primer_bind     7435..7456
                     /label=bGH-Blst(2).F
     primer_bind     7435..7453
                     /label=bGH-Blst(3).F
     primer_bind     7435..7453
                     /label=bGH-Blst(4).F
     primer_bind     7440..7458
                     /label=pU3-NeoR.F
     polyA_signal    7441..7665
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7441..7461
                     /label=pUtopia53.HiR
     primer_bind     complement(7462..7485)
                     /label=TG1.6.10
     primer_bind     7462..7485
                     /label=TG1.6.9
     primer_bind     complement(7498..7512)
                     /label=AAVS1.DxR
                     /note="AAVS1 diagnostic PCR, forward. PCR product 394bp for
                     wild-type (NC_000019.10). Products on potentially 
                     unintended templates:>NC_000001.11 Homo sapiens chromosome 
                     1, GRCh38.p7 Primary Assemblyproduct length = 2209Features 
                     associated with this product:  T-cell surface glycoprotein 
                     CD3 zeta chain isoform 2 prec...  T-cell surface 
                     glycoprotein CD3 zeta chain isoform 1 prec... product 
                     length = 3051Features associated with this product: group 
                     IIF secretory phospholipase A2 isoform X1 group IIF 
                     secretory phospholipase A2>NC_000004.12 Homo sapiens 
                     chromosome 4, GRCh38.p7 Primary Assembly product length = 
                     3333 Features associated with this product: calcium 
                     uniporter regulatory subunit MCUb, mitochondrial calcium 
                     uniporter regulatory subunit MCUb, mitochondrial 
                     >NC_000007.14 Homo sapiens chromosome 7, GRCh38.p7 Primary 
                     Assembly  product length = 1258 Features flanking this 
                     product: 104529 bp at 5' side: histone-lysine 
                     N-methyltransferase EZH2 isoform X22 47538 bp at 3' side: 
                     protein disulfide-isomerase A4 precursor"
     primer_bind     7612..7636
                     /label=pPT-mLin-target-general.HiR
     primer_bind     complement(7612..7636)
                     /label=pPT-mLin-scaffold-general.HiF
     primer_bind     complement(7616..7636)
                     /label=pRIME-mLin-GFP-1.HiR
     primer_bind     7616..7636
                     /label=pRIME-mLin-GFP-2.HiF
     primer_bind     complement(7616..7635)
                     /label=pRIME-mLin-Twist-1.HiR
     primer_bind     complement(7616..7635)
                     /label=proL-NF-GFPv4-1.HiR
     primer_bind     7616..7635
                     /label=proL-NF-GFPv4-2.HiR
     primer_bind     complement(7645..7665)
                     /label=bGH-pA-r
     primer_bind     complement(7646..7665)
                     /label=pPT-mLin-target.HiR
     primer_bind     7646..7665
                     /label=pRIME-mLin-Twist-1.HiR
     primer_bind     7646..7665
                     /label=pPT-mLin-scaffold.HiF
     primer_bind     complement(7646..7665)
                     /label=pPT-mLin-Backbone-2.HiR
     primer_bind     complement(7649..7667)
                     /label=RFP-AAVS1.HiR
     primer_bind     complement(7660..7679)
                     /label=pHR-mLin-GFP-3.HiR
     primer_bind     7661..7680
                     /label=proL-NF-GFPv4-1.HiR
     primer_bind     complement(7661..7680)
                     /label=proL-NF-GFPv4-2.HiR
     primer_bind     7661..7680
                     /label=pUT-mPF-GFP-1.HiF
     primer_bind     7677..7691
                     /label=pPT2-backbone-2_v2.HiR
     primer_bind     7678..7693
                     /label=pPT2-Backbone-2.HiR
     primer_bind     7741..7786
                     /label=Blst-FRT-Ori.F
     primer_bind     complement(7747..7766)
                     /label=pPT-mLin-target-general.HiR
     primer_bind     complement(7747..7766)
                     /label=pPT-mLin-Split-2.HiR
     primer_bind     7747..7766
                     /label=pPT-mLin-Split-1.HiR
     primer_bind     7747..7766
                     /label=pPT-mLin-scaffold-general.HiF
     primer_bind     7750..7769
                     /label=CRISPRa-donor-vector.HiF
     primer_bind     7750..7769
                     /label=put_NF_BACH1_P2A_GFP-1.HiF
     primer_bind     7755..7775
                     /label=pUT-mPF-GFP-1.HiF
     primer_bind     7761..7786
                     /label=Blst-FRT-OL-f
     primer_bind     7772..7795
                     /label=GFP_Linearizer_FRT.HiF
     polyA_site      7800..8194
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     primer_bind     complement(7826..7852)
                     /label=B-globin pA end outward
     primer_bind     complement(8182..8202)
                     /label=pPT-mLin-target_v2.HiR
     primer_bind     8182..8202
                     /label=pPT-mLin-scaffold_v3.HiF
     primer_bind     complement(8182..8202)
                     /label=pPT-mLin-target_v3.HiR
     gene            complement(8202..8625)
                     /product="Blasticidin Resistance gene"
                     /label=BsrS2
     primer_bind     8351..8370
                     /label=NF-BACH1 tail internal V1
     primer_bind     8396..8420
                     /label=NF-BACH1 tail internal V2
     primer_bind     complement(8601..8625)
                     /label=pPT-mPF-GFP-BACH1-2.HiR
     primer_bind     complement(8606..8645)
                     /label=PPT-to-PUT change-1.HiF
     primer_bind     8606..8645
                     /label=PPT-to-PUT change-2.HiR
     primer_bind     8606..8645
                     /label=PPT-to-PUT change-2.HiR
     primer_bind     8606..8645
                     /label=PPT-to-PUT change-2.HiR (3)
     primer_bind     8612..8628
                     /label=nvn.1.15
     primer_bind     8614..8645
                     /label=PPT-to-PUT change-2.HiR
     primer_bind     8614..8645
                     /label=PPT-to-PUT change-2.HiR
     primer_bind     8614..8645
                     /label=PPT-to-PUT change-2.HiR (2)
     primer_bind     8614..8645
                     /label=PPT-to-PUT change-2.HiR (2) (2)
     primer_bind     8625..8645
                     /label=pPT2-Cas13-EGFP-KD-test-1.HiR
     primer_bind     8626..8645
                     /label=RIME-1.1
     primer_bind     8626..8645
                     /label=RIME-1.1-2
     promoter        complement(8634..8931)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     8634..8653
                     /label=RIME-2-1.HiFv2
     primer_bind     8938..8977
                     /label=PPT-to-PUT change-1.HiR
     primer_bind     complement(8938..8977)
                     /label=PPT-to-PUT change-2.HiF
     primer_bind     complement(8938..8959)
                     /label=pPT2-Cas13-EGFP-KD-test-1.HiF
     misc_feature    8958..8976
                     /label=SA
     misc_feature    complement(8977..9010)
                     /label=F3
     primer_bind     complement(8977..9010)
                     /label=pHR-mLin-GFP-3.HiR
     primer_bind     8977..8991
                     /label=pPT2-Backbone-2.HiR
     primer_bind     8977..8990
                     /label=nvn.1.15
     intron          9011..9054
                     /label=HA1-AAVS1
     misc_feature    9017..9054
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(9017..9034)
                     /label=AAVS1-HA-Left-C-Term-Seq-f
     primer_bind     9017..9034
                     /label=AAVS1-HA-Left-C-Term-Seq-r
     primer_bind     9027..9048
                     /label=protrap_Fluo-AAVS1_ext.HiF
     primer_bind     9035..9074
                     /label=pRIME-mLin-Twist-2.HiR
     primer_bind     complement(9035..9074)
                     /label=pRIME-mLin-Twist-1.HiF
     primer_bind     9054..9074
                     /label=pPT2-Backbone-1.HiF
     primer_bind     9055..9074
                     /label=nvn.1.17
     primer_bind     complement(9055..9074)
                     /label=nvn.1.20
     primer_bind     9073..9092
                     /label=pUT-AmpR-addon-2.HiF
     promoter        9077..9181
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9126..9145
                     /label=AmpPromoter-split.fwd
     primer_bind     complement(9126..9145)
                     /label=AmpPromoter-split.rev
     CDS             join(9182..9250,9251..9903,9904..10042)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9660..9679)
                     /label=AmpR split fwd
     primer_bind     9660..9679
                     /label=AmpR-split.rev
                     /note="Primer to split AmpR for HiFi assembly"
     primer_bind     10040..10079
                     /label=pPT-mLin-BB_v2-2.HiF
     primer_bind     complement(10040..10079)
                     /label=pPT-mLin-BB_v2-1.HiR
     primer_bind     complement(10043..10062)
                     /label=nvn.1.18
     primer_bind     10043..10059
                     /label=nvn.1.19
     primer_bind     complement(10045..10062)
                     /label=Ext_HiF
     primer_bind     complement(10045..10062)
                     /label=pRIME-mLin-GFP-1.HiF
     primer_bind     10045..10059
                     /label=pRIME-mLin-GFP-2.HiR
     primer_bind     10060..10085
                     /label=KRASB-HSV.F
     primer_bind     complement(10060..10078)
                     /label=nvn.1.17
     primer_bind     10060..10078
                     /label=nvn.1.20
     primer_bind     complement(10062..10081)
                     /label=pUT-AmpR-addon-2.HiR
     promoter        10075..10571
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(10440..10462)
                     /label=TG1.16.4
     5'UTR           10613..10652
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     primer_bind     10615..10633
                     /label=HSVTK_UTR_seq_R-r
     primer_bind     complement(10656..10675)
                     /label=noBach1 Proto Frag1 Rev
     primer_bind     10664..10683
                     /label=noBach1_FragB-F_identical-overlap_V1
     primer_bind     complement(10664..10683)
                     /label=noBach1_FragA-R_identical-overlap_V1
     primer_bind     complement(10664..10683)
                     /label=nvn1.1.2
     primer_bind     10664..10683
                     /label=nvn1.1.3
     CDS             10672..11802
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /translation="MASYPCHQHASAFDQAARSRGHNNRRTALRPRRQQKATEVRLEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGAAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     primer_bind     10676..10695
                     /label=noBach1 Proto Frag2 Forward
     primer_bind     10948..10967
                     /label=HSVTKfwd
     primer_bind     11124..11136
                     /label=TG1.6.9
     primer_bind     11225..11240
                     /label=FRT-Ori-OL-f
     primer_bind     complement(11372..11391)
                     /label=noBach1 Mutagen Split Frag1
     primer_bind     complement(11438..11449)
                     /label=pPT-mLin-scaffold-general.HiF
     primer_bind     complement(11633..11652)
                     /label=no Bach1 mutagen split frag2
     primer_bind     complement(11785..11804)
                     /label=noBach1 Proto FragB Forw
     primer_bind     11805..11824
                     /label=noBach1 Proto FragA rev
     polyA_signal    11844..11892
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(11874..11893)
                     /label=HSV_TK-FRT-Rev
     primer_bind     11879..11900
                     /label=CodA-FRT3(2).F
     primer_bind     complement(join(11882..11901,1..20))
                     /label=pPT-mLin-BB_v2-2.HiR
     primer_bind     join(11882..11901,1..20)
                     /label=pPT-mLin-BB_v2-1.HiF
     primer_bind     complement(11882..11901)
                     /label=KRASB-HSV.R
ORIGIN
        1 ctcataggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
       61 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
      121 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
      181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
      241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
      421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      601 aaaaacgccg aagttcctat actttctaga gaataggaac ttcacgggcc agatatacgc
      661 gacgggccag atatacgcga cgggccagat atacgcgacg ggccagatat acgcgacggg
      721 ccagatatac gcgacgggcc agatatacgc gacgggccag atatacgcga cgggccagat
      781 atacgcgacg ggccagatat acgcgacggg ccagatatac gcgacgggcc agatatacgc
      841 gacgggccag atatacgcga cgggccagat atacgcgttg acattgatta ttgactagtt
      901 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      961 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     1021 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     1081 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     1141 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     1201 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     1261 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     1321 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     1381 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     1441 gggaggtcta tataagcatc cctatcagtg atagagatca gatctcccta tcagtgatag
     1501 agagctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg actctagcgt
     1561 ttaaacttaa gcttggtacc cggggatcct ctagggcctc tgagctattc cagaagtagt
     1621 gaagaggctt ttttggaggc ctaggctttt gcaaaaagct ccggatcgat cctgagaact
     1681 tcagggtgag tttggggacc cttgattgtt ctttcttttt cgctattgta aaattcatgt
     1741 tatatggagg gggcaaagtt ttcagggtgt tgtttagaat gggaagatgt cccttgtatc
     1801 accatggacc ctcatgataa ttttgtttct ttcactttct actctgttga caaccattgt
     1861 ctcctcttat tttcttttca ttttctgtaa ctttttcgtt aaactttagc ttgcatttgt
     1921 aacgaatttt taaattcact tttgtttatt tgtcagattg taagtacttt ctctaatcac
     1981 ttttttttca aggcaatcag ggtatattat attgtacttc agcacagttt tagagaacaa
     2041 ttgttataat taaatgataa ggtagaatat ttctgcatat aaattctggc tggcgtggaa
     2101 atattcttat tggtagaaac aactacatcc tggtcatcat cctgcctttc tctttatggt
     2161 tacaatgata tacactgttt gagatgagga taaaatactc tgagtccaaa ccgggcccct
     2221 ctgctaacca tgttcatgcc ttcttctttt tcctacaggt cctgcaggcg ccaccatggg
     2281 ttctagactg gacaagagca aagtcataaa ctctgctctg gaattactca atgaagtcgg
     2341 tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc agcctaccct
     2401 gtactggcac gtgaagaaca agcgggccct gctcgatgcc ctggcaatcg agatgctgga
     2461 caggcatcat acccacttct gccccctgga aggcgagtca tggcaagact ttctgcggaa
     2521 caacgccaag tcattccgct gtgctctcct ctcacatcgc gacggggcta aagtgcatct
     2581 cggcacccgc ccaacagaga aacagtacga aaccctggaa aatcagctcg cgttcctgtg
     2641 tcagcaaggc ttctccctgg agaacgcact gtacgctctg tccgccgtgg gccactttac
     2701 actgggctgc gtattggagg atcaggagca tcaagtagca aaagaggaaa gagagacacc
     2761 taccaccgat tctatgcccc cacttctgag acaagcaatt gagctgttcg accatcaggg
     2821 agccgaacct gccttccttt tcggcctgga actaatcata tgtggcctgg agaaacagct
     2881 aaagtgcgaa agcggcgggc caaaaaagaa gagaaaggga agcggagcta ctaacttcag
     2941 cctgctgaag caggctggag acgtggagga gaaccctgga cctatggtga gcaagggcga
     3001 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     3061 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     3121 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     3181 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     3241 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     3301 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     3361 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     3421 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     3481 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     3541 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     3601 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     3661 ctccaccggc ggcatggacg agctgtacaa gggcagcgga gctactaact tcagcctgct
     3721 gaagcaggct ggagacgtgg aggagaaccc tggacccggg atgatcgaga aaaagaagtc
     3781 cttcgctaag ggcatgggcg tgaagtccac actcgtgtcc ggctccaaag tgtacatgac
     3841 aaccttcgcc gaaggcagcg acgccaggct ggaaaagatc gtggagggcg acagcatcag
     3901 gagcgtgaat gagggcgagg ccttcagcgc tgaaatggcc gataaaaacg ccggctataa
     3961 gatcggcaac gccaaattca gccatcctaa gggctacgcc gtggtggcta acaaccctct
     4021 gtatacagga cccgtccagc aggatatgct cggcctgaag gaaactctgg aaaagaggta
     4081 cttcggcgag agcgctgatg gcaatgacaa tatttgtatc caggtgatcc ataacatcct
     4141 ggacattgaa aaaatcctcg ccgaatacat taccaacgcc gcctacgccg tcaacaatat
     4201 ctccggcctg gataaggaca ttattggatt cggcaagttc tccacagtgt atacctacga
     4261 cgaattcaaa gaccccgagc accatagggc cgctttcaac aataacgata agctcatcaa
     4321 cgccatcaag gcccagtatg acgagttcga caacttcctc gataacccca gactcggcta
     4381 tttcggccag gcctttttca gcaaggaggg cagaaattac atcatcaatt acggcaacga
     4441 atgctatgac attctggccc tcctgagcgg actgagacac tgggtggtcc acaacaacga
     4501 agaagagtcc aggatctcca ggacctggct ctacaacctc gataagaacc tcgacaacga
     4561 atacatctcc accctcaact acctctacga caggatcacc aatgagctga ccaactcctt
     4621 ctccaagaac tccgccgcca acgtgaacta tattgccgaa actctgggaa tcaaccctgc
     4681 cgaattcgcc gaacaatatt tcagattcag cattatgaaa gagcagaaaa acctcggatt
     4741 caatatcacc aagctcaggg aagtgatgct ggacaggaag gatatgtccg agatcaggaa
     4801 aaatcataag gtgttcgact ccatcaggac caaggtctac accatgatgg actttgtgat
     4861 ttataggtat tacatcgaag aggatgccaa ggtggctgcc gccaataagt ccctccccga
     4921 taatgagaag tccctgagcg agaaggatat ctttgtgatt aacctgaggg gctccttcaa
     4981 cgacgaccag aaggatgccc tctactacga tgaagctaat agaatttgga gaaagctcga
     5041 aaatatcatg cacaacatca aggaatttag gggaaacaag acaagagagt ataagaagaa
     5101 ggacgcccct agactgccca gaatcctgcc cgctggccgt gatgtttccg ccttcagcaa
     5161 actcatgtat gccctgacaa tgttcctgga tggcaaggag atcaacgacc tcctgaccac
     5221 cctgattaat aaattcgata acatccagag cttcctgaag gtgatgcctc tcatcggagt
     5281 caacgctaag ttcgtggagg aatacgcctt tttcaaagac tccgccaaga tcgccgatga
     5341 gctgaggctg atcaagtcct tcgctagaat gggagaacct attgccgatg ccaggagggc
     5401 catgtatatc gacgccatcc gtattttagg aaccaacctg tcctatgatg agctcaaggc
     5461 cctcgccgac accttttccc tggacgagaa cggaaacaag ctcaagaaag gcaagcacgg
     5521 catgagaaat ttcattatta ataacgtgat cagcaataaa aggttccact acctgatcag
     5581 atacggtgat cctgcccacc tccatgagat cgccaaaaac gaggccgtgg tgaagttcgt
     5641 gctcggcagg atcgctgaca tccagaaaaa acagggccag aacggcaaga accagatcga
     5701 caggtactac gaaacttgta tcggaaagga taagggcaag agcgtgagcg aaaaggtgga
     5761 cgctctcaca aagatcatca ccggaatgaa ctacgaccaa ttcgacaaga aaaggagcgt
     5821 cattgaggac accggcaggg aaaacgccga gagggagaag tttaaaaaga tcatcagcct
     5881 gtacctcacc gtgatctacc acatcctcaa gaatattgtc aatatcaacg ccaggtacgt
     5941 catcggattc cattgcgtcg agcgtgatgc tcaactgtac aaggagaaag gctacgacat
     6001 caatctcaag aaactggaag agaagggatt cagctccgtc accaagctct gcgctggcat
     6061 tgatgaaact gcccccgata agagaaagga cgtggaaaag gagatggctg aaagagccaa
     6121 ggagagcatt gacagcctcg agagcgccaa ccccaagctg tatgccaatt acatcaaata
     6181 cagcgacgag aagaaagccg aggagttcac caggcagatt aacagggaga aggccaaaac
     6241 cgccctgaac gcctacctga ggaacaccaa gtggaatgtg atcatcaggg aggacctcct
     6301 gagaattgac aacaagacat gtaccctgtt caggaacaag gccgtccatc tggaagtggc
     6361 caggtatgtc cacgcctata tcaacgacat tgccgaggtc aattcctact tccaactgta
     6421 ccattacatc atgcagagaa ttatcatgaa tgagaggtac gagaaaagca gcggaaaggt
     6481 gtccgagtac ttcgacgctg tgaatgacga gaagaagtac aacgataggc tcctgaaact
     6541 gctgtgtgtg cctttcggct actgtatccc caggtttaag aacctgagca tcgaggccct
     6601 gttcgatagg aacgaggccg ccaagttcga caaggagaaa aagaaagtgt ccggcaattc
     6661 cactagtgct cccaagaaaa agcgcaaggt aggtggaagc ccagcagcta aaagagttaa
     6721 attggatgga tccgattata aggatcacga tggagattac aaggaccacg acatagacta
     6781 caaagatgat gacgacaagt aacgtacgta agaattcgat tcgtcagtag ggttgtaaag
     6841 gtttttcttt tcctgagaaa acaacctttt gttttctcag gttttgcttt ttggcctttc
     6901 cctagcttta aaaaaaaaaa agcaaaactc accgaggcag ttccatagga tggcaaggcc
     6961 gcgactctag agtcgacctg caggcatgca agcttgatat caggccgcaa tcaacctctg
     7021 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     7081 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     7141 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     7201 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     7261 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     7321 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     7381 aattccgtgg tgttgtcctc gagtctagag ggcccgttta aacccgctga tcagcctcga
     7441 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     7501 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     7561 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     7621 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     7681 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     7741 cgggtgtggt ggttacgcgc taaagcaagt aaaacctcta caaatgtggt atggaattct
     7801 aaaatacagc atagcaaaac tttaacctcc aaatcaagcc tctacttgaa tccttttctg
     7861 agggatgaat aaggcatagg catcaggggc tgttgccaat gtgcattagc tgtttgcagc
     7921 ctcaccttct ttcatggagt ttaagatata gtgtattttc ccaaggtttg aactagctct
     7981 tcatttcttt atgttttaaa tgcactgacc tcccacattc cctttttagt aaaatattca
     8041 gaaataattt aaatacatca ttgcaatgaa aataaatgtt ttttattagg cagaatccag
     8101 atgctcaagg cccttcataa tatcccccag tttagtagtt ggacttaggg aacaaaggaa
     8161 cctttaatag aaattggaca gcaagaaagc gagcttctag ctttagttcc tggtgtactt
     8221 gagggggatg agttcctcaa tggtggtttt gaccagcttg ccattcatct caatgagcac
     8281 aaagcagtca ggagcatagt cagagatgag ctctctgcac atgccacagg ggctgaccac
     8341 cctgatggat ctgtccacct catcagagta ggggtgcctg acagccacaa tggtgtcaaa
     8401 gtccttctgc ccgttgctca cagcagaccc aatggcaatg gcttcagcac agacagtgac
     8461 cctgccaatg taggcctcaa tgtggacagc agagatgatc tccccagtct tggtcctgat
     8521 ggccgccccg acatggtgct tgttgtcctc atagagcatg gtgatcttct cagtggcgac
     8581 ctccaccagc tccagatcct gctgagagat gttgaaggtc ttcatggtag cctccaaaaa
     8641 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa
     8701 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg
     8761 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca
     8821 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat
     8881 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac
     8941 acacattcca cagctgcgaa gagaagaggt caggccgaag ttcctatact atttgaagaa
     9001 taggaacttc aagcttgccc cactgtgggg tggaggggac agataaaagt accctggcac
     9061 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     9121 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     9181 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     9241 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     9301 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     9361 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     9421 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     9481 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     9541 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     9601 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     9661 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     9721 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     9781 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     9841 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     9901 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     9961 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
    10021 ctcactgatt aagcattggt aactgtcaga ccaagtttat tctacggttc taccgggtag
    10081 gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt
    10141 ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac
    10201 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt
    10261 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca
    10321 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca
    10381 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
    10441 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag
    10501 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat
    10561 ctccgggcct ttcgacctgc agcgacccgc taacagcgtc aacagcgtgc cgcagatctt
    10621 ggtggcgtga aactcccgca cctcttcggc cagcgccttg tagaagcgcg tatggcttcg
    10681 tacccctgcc atcaacacgc gtctgcgttc gaccaggctg cgcgttctcg cggccataac
    10741 aaccgacgta cggcgttgcg ccctcgccgg caacaaaaag ccacggaagt ccgcctggag
    10801 cagaaaatgc ccacgctact gcgggtttat atagacggtc cccacgggat ggggaaaacc
    10861 accaccacgc aactgctggt ggccctgggt tcgcgcgacg atatcgtcta cgtacccgag
    10921 ccgatgactt actggcgggt gttgggggct tccgagacaa tcgcgaacat ctacaccaca
    10981 caacaccgcc tcgaccaggg tgagatatcg gccggggacg cggcggtggt aatgacaagc
    11041 gcccagataa caatgggcat gccttatgcc gtgaccgacg ccgttctggc tcctcatatc
    11101 gggggggagg ctgggagctc acatgccccg cccccggccc tcaccctcat cttcgaccgc
    11161 catcccatcg ccgccctcct gtgctacccg gccgcgcgat accttatggg cagcatgacc
    11221 ccccaggccg tgctggcgtt cgtggccctc atcccgccga ccttgcccgg cacaaacatc
    11281 gtgttggggg cccttccgga ggacagacac atcgaccgcc tggccaaacg ccagcgcccc
    11341 ggcgagcggc ttgacctggc tatgctggcc gcgattcgcc gcgtttatgg gctgcttgcc
    11401 aatacggtgc ggtatctgca gggcggcggg tcgtggcggg aggattgggg acagctttcg
    11461 ggggcggccg tgccgcccca gggtgccgag ccccagagca acgcgggccc acgaccccat
    11521 atcggggaca cgttatttac cctgtttcgg gcccccgagt tgctggcccc caacggcgac
    11581 ctgtacaacg tgtttgcctg ggccttggac gtcttggcca aacgcctccg tcccatgcac
    11641 gtctttatcc tggattacga ccaatcgccc gccggctgcc gggacgccct gctgcaactt
    11701 acctccggga tggtccagac ccacgtcacc acccccggct ccataccgac gatctgcgac
    11761 ctggcgcgca cgtttgcccg ggagatgggg gaggctaact gaaacacgga aggagacaat
    11821 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg
    11881 ggtcgtttgt tcataaacgc g
//
