LOCUS       Exported               16802 bp ds-DNA     circular SYN 20-OCT-2020
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 16802)
  AUTHORS   yy
  TITLE     Direct Submission
  JOURNAL   Exported Apr 21, 2022 from SnapGene 4.2.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..16802
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    1..2341
                     /label=FAST-RED
                     /note="color: #a6acb3"
     primer_bind     2358..2379
                     /label=BsaI_GGAG_Pro_F
                     /note="color: black; sequence: 
                     AAGGTCTCAGGAGGAACCGCAACGTTGAAGG"
     promoter        2362..2540
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        2394..2576
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     2523..2543
                     /label=Nos_promoter_F
                     /note="color: black; sequence: CCCCTCGGTATCCAATTAGAG"
     terminator      3176..3428
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
                     /note="color: #ffffff"
     primer_bind     complement(3362..3382)
                     /label=Nos_Ter_R
                     /note="color: black; sequence: TGCGCGCTATATTTTGTTTTC"
     primer_bind     complement(3404..3432)
                     /label=BsaI_CGCT_Ter_R
                     /note="color: black; sequence: 
                     AAGGTCTCAAGCGGATCTAGTAACATAGATGACACCGC"
     misc_feature    3453..4087
                     /label=E9 terminator
                     /note="color: #a6acb3"
     primer_bind     3999..4020
                     /label=rbcs check R
                     /note="color: black; sequence: CTCAGTAGGATTCTGGTGTGTG"
     primer_bind     4036..4057
                     /label=E9_ter_R
                     /note="color: black; sequence: GCATTGAACTTGACGAACGTTG"
     primer_bind     complement(4036..4054)
                     /label=pRW sequencing F2
                     /note="color: black; sequence: CGTTCGTCAAGTTCAATGC"
     primer_bind     4088..4112
                     /label=066_REV
                     /note="intro stop codon"
                     /note="color: black; sequence: 
                     atGAAGACaaAGCGTTAgccacctccaaccttcct"
     misc_feature    complement(4095..4103)
                     /label=GGGS-PmCDA1-UGI-NLS
                     /note="GGGS-PmCDA1-UGI-NLS"
                     /note="color: #993366; direction: LEFT"
     misc_feature    complement(4104..8240)
                     /label=SpCas9-NG(D10A)-NLS
                     /note="color: #ff0000; direction: LEFT"
     CDS             complement(4104..4124)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     misc_feature    complement(6522..8240)
                     /label=Cas9-NG (2 base different)
                     /note="N-terminal portion of Streptococcus pyogenes Cas9 
                     (Zetsche et al., 2015)"
                     /note="color: #993366; direction: LEFT"
     primer_bind     complement(8214..8241)
                     /label=066_FOR
                     /note="color: black; sequence: 
                     atGAAGACctaatggataagaagtactctatcggactc"
     primer_bind     8238..8256
                     /label=add in 066_R
                     /note="color: black; sequence: 
                     atGAAGACaaCATTCTGCCGCCAGAGCTG"
     CDS             complement(8242..9431)
                     /codon_start=1
                     /label=TadA_Hv
                     /note="color: #993366"
                     /translation="MSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGW
                     NRPIGRHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRVVF
                     GARDAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQ
                     SSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSSEVEFSHEYWMRHALTLAKRARD
                     EREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVT
                     FEPCVMCAGAMIHSRIGRVVFGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECA
                     ALLCYFFRMPRQVFNAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGG"
     misc_feature    8242..8639
                     /label=B27r
                     /label=nonstandard type: Seq.
                     /note="color: #a6acb3"
     primer_bind     8834..8852
                     /label=Linker_moclo_R
                     /note="color: black; sequence: AAGGTCTCAAGATCCGCCGCTAGACCC"
     misc_feature    complement(9133..9432)
                     /label=L2r
                     /label=nonstandard type: Seq.
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    9433..11124
                     /label=RPS5a promoter
                     /note="color: #a6acb3"
     primer_bind     complement(9492..9511)
                     /label=RPS5A Sequence F
                     /note="color: black; sequence: CTTTCTCCAAATCCTCTCTC"
     primer_bind     complement(9519..9538)
                     /label=RPS5A Sequence F2
                     /note="color: black; sequence: CGACGCAAACGATCTTCAGG"
     primer_bind     10232..10251
                     /label=AtRPS5A_promoter_R
                     /note="color: black; sequence: CACTCCATTGATGTTCCGTG"
     misc_feature    11145..11346
                     /label=pU6-26
                     /note="color: #a6acb3"
     primer_bind     11207..11225
                     /label=U6 promoter sgRNA sequencing_2
                     /note="color: black; sequence: AAGCAGGCCCATTTATATG"
     primer_bind     11253..11272
                     /label=U6 promoter sgRNA sequencing
                     /note="color: black; sequence: GGCCCATTTAAGTTGAAAAC"
     primer_bind     complement(11328..11357)
                     /label=BsaI_p4_F
                     /note="color: black; sequence: 
                     aaGGTCTCaCAATcactacttcgactctagct"
     primer_bind     11347..11372
                     /label=addLacZ_F
                     /note="color: black; sequence: 
                     ATGAAGACAAATTGTGAGACCGTCACAGCTTGTCTG"
     CDS             complement(11447..11770)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /note="color: #993366"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     11452..11471
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
                     /note="color: black; sequence: CGGCATCAGAGCAGATTGTA"
     primer_bind     11628..11647
                     /label=pMLBart Seq-F
                     /note="color: black; sequence: GCTGCAAGGCGATTAAGTTG"
     primer_bind     11660..11679
                     /label=Fw_NC
                     /note="color: black; sequence: GTTTTCCCAGTCACGACGTT"
     primer_bind     11665..11687
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
                     /note="color: black; sequence: CCCAGTCACGACGTTGTAAAACG"
     primer_bind     complement(11679..11696)
                     /label=M13-F-Reverse Complement
                     /note="color: black; sequence: ACTGGCCGTCGTTTTACA"
     primer_bind     11679..11696
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
                     /note="color: black; sequence: TGTAAAACGACGGCCAGT"
     primer_bind     11680..11697
                     /label=FW-1-K153A: M13 fwd
                     /note="color: black; sequence: GTAAAACGACGGCCAGTG"
     primer_bind     11680..11696
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     misc_feature    11697..11753
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
                     /note="color: #99ccff"
     primer_bind     complement(11766..11782)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     11766..11782
                     /label=M13-R-Reverse Complement
                     /note="color: black; sequence: GTCATAGCTGTTTCCTG"
     primer_bind     complement(11766..11782)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
                     /note="color: black; sequence: CAGGAAACAGCTATGAC"
     primer_bind     complement(11779..11801)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
                     /note="color: black; sequence: AGCGGATAACAATTTCACACAGG"
     protein_bind    11790..11806
                     /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"
     promoter        complement(11814..11844)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 11814 .. 11820 / #ffffff / -10
                        2: 11821 .. 11838 / #ffffff
                        3: 11839 .. 11844 / #ffffff / -35"
     primer_bind     complement(11814..11832)
                     /label=pMLBart Seq-R
                     /note="color: black; sequence: GCTTCCGGCTCGTATGTTG"
     primer_bind     complement(11817..11837)
                     /label=pGEMTEasy insert_R
                     /note="color: black; sequence: TTTATGCTTCCGGCTCGTATG"
     protein_bind    11859..11880
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     primer_bind     complement(11927..11946)
                     /label=addLacZ_R
                     /note="color: black; sequence: 
                     ATGAAGACAAAAACTGAGACCGCAGCTGGC"
     primer_bind     11936..11965
                     /label=BsaI_p4_R
                     /note="color: black; sequence: 
                     aaGGTCTCaGTTTAAGAGCTATGCTGGAAACA"
     primer_bind     11942..11962
                     /label=AarI_add
                     /note="color: black; sequence: 
                     tgtggtctcaATTGaaatgcaggtgatgactcacctgcataaGTTTAAGAGCTATGCTG
                     GAA"
     primer_bind     11943..11962
                     /label=pguide1_FOR
                     /note="color: black; sequence: 
                     TGTGGTCTCAATTGGGAATTGGTAAGACTACCATGTTTAAGAGCTATGCTGGAA"
     primer_bind     11943..11962
                     /label=pguide2_FOR
                     /note="color: black; sequence: 
                     TGTGGTCTCAATTGGGTAAGACTACCATTGCTAGGTTTAAGAGCTATGCTGGAA"
     primer_bind     11943..11962
                     /label=R422Gguide_FOR
                     /note="color: black; sequence: 
                     TGTGGTCTCAATTGAGTGAGATATGAAAGATTGTGTTTAAGAGCTATGCTGGAA"
     primer_bind     11989..12014
                     /label=pICH47751_sgRNA_check_F
                     /note="color: black; sequence: AGTCCGTTATCAACTTGAAAAAGTGG"
     primer_bind     complement(12080..12099)
                     /label=sgRNA_R
                     /note="color: black; sequence: 
                     tgtggtctcaAGCGACCCCAGAAATTGAAC"
     primer_bind     complement(12152..12176)
                     /label=pICH47751_check_F
                     /note="color: black; sequence: CGCCAATATATCCTGTCAAACACTG"
     misc_feature    12159..12183
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
                     /note="color: #ffe4c4"
     primer_bind     complement(12162..12184)
                     /label=MoClo_check_F
                     /note="color: black; sequence: GGTTTACCCGCCAATATATCCTG"
     CDS             12463..13092
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /note="color: #993366"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             13521..14594
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /note="color: #993366"
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     primer_bind     14027..14041
                     /label=pICH47751_check_R
                     /note="color: black; sequence: AACTTAATAACACATTGCGGACGT"
     rep_origin      14660..14854
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /note="color: #ffff00"
     promoter        14916..15020
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             15021..15815
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /note="color: #ccffcc"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     primer_bind     15818..15852
                     /label=HR4 ICE106 R
                     /note="color: black; sequence: 
                     GTCAGACCAAGTTTACTCATATATACTTTAGATTG"
     rep_origin      15986..16574
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     16475..16494
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     primer_bind     16479..16500
                     /label=pYYL001_Seq_R_1
                     /note="color: black; sequence: AACGCCTGGTATCTTTATAGTC"
     primer_bind     16651..16674
                     /label=MoClo_check_R
                     /note="color: black; sequence: CCTGTATCGAGTGGTGATTTTGTG"
     primer_bind     16651..16674
                     /label=pICH47751_sgRNA_check_R
                     /note="color: black; sequence: CCTGTATCGAGTGGTGATTTTGTG"
     misc_feature    16710..16734
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
                     /note="color: #ffe4c4"
     primer_bind     16751..16774
                     /label=pICH47751_check_R
                     /note="color: black; sequence: AACTTAATAACACATTGCGGACGT"
ORIGIN
        1 tgccgaattc ggatccggag ttctagaatg tcgcggaaca aattttaaaa ctaaatccta
       61 aatttttcta attttgttgc caatagtgga tatgtgggcc gtatagaagg aatctattga
      121 aggcccaaac ccatactgac gagcccaaag gttcgttttg cgttttatgt ttcggttcga
      181 tgccaacgcc acattctgag ctaggcaaaa aacaaacgtg tctttgaata gactcctctc
      241 gttaacacat gcagcggctg catggtgacg ccattaacac gtggcctaca attgcatgat
      301 gtctccattg acacgtgact tctcgtctcc tttcttaata tatctaacaa acactcctac
      361 ctcttccaaa atatatacac atctttttga tcaatctctc attcaaaatc tcattctctc
      421 tagtaaacaa gaacaaaaaa atggcggata cagctagagg aacccatcac gatatcatcg
      481 gcagagatca gtacccgatg atgggccgag atcgtgacca gtaccagatg tccggacgag
      541 gatctgacta ctccaagtct aggcagattg ctaaagctgc aactgctgtc acagctggtg
      601 gttccctcct tgttctctcc agccttaccc ttgttggaac tgtcatagct ttgactgttg
      661 caacacctct gctcgttatc ttcagcccaa tccttgtccc ggctctcatc acagttgcac
      721 tcctcatcac cggttttctt tcctctggag ggtttggcat tgccgctata accgttttct
      781 cttggattta caagtaagca cacatttatc atcttacttc ataattttgt gcaatatgtg
      841 catgcatgtg ttgagccagt agctttggat caattttttt ggtcgaataa caaatgtaac
      901 aataagaaat tgcaaattct agggaacatt tggttaacta aatacgaaat ttgacctagc
      961 tagcttgaat gtgtctgtgt atatcatcta tataggtaaa atgcttggta tgatacctat
     1021 tgattgtgaa taggtacgca acgggagagc acccacaggg atcagacaag ttggacagtg
     1081 caaggatgaa gttgggaagc aaagctcagg atctgaaaga cagagctcag tactacggac
     1141 agcaacatac tggtggggaa catgaccgtg accgtactcg tggtggccag cacactacta
     1201 tgagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtgaaca
     1261 accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
     1321 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca
     1381 gcttcatgta cggcagcaga accttcatca accacaccca gggcatcccc gacttcttta
     1441 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gatgggggcg
     1501 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
     1561 tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     1621 aggccaacac cgagatgctg taccccgctg acggcggcct ggaaggcaga agcgacatgg
     1681 ccctgaagct cgtgggcggg ggccacctga tctgcaactt caagaccaca tacagatcca
     1741 agaaacccgc taagaacctc aagatgcccg gcgtctacta tgtggaccac agactggaaa
     1801 gaatcaagga ggccgacaaa gaaacctacg tcgagcagca cgaggtggct gtggccagat
     1861 actgcgacct ccctagcaaa ctggggcaca agtgagctta ccccactgat gtcatcgtca
     1921 tagtccaata actccaatgt cggggagtta gtttatgagg aataaagtgt ttagaatttg
     1981 atcaggggga gataataaaa gccgagtttg aatctttttg ttataagtaa tgtttatgtg
     2041 tgtttctata tgttgtcaaa tggtaccatg ttttttttcc tctctttttg taacttgcaa
     2101 gtgttgtgtt gtactttatt tggcttcttt gtaagttggt aacggtggtc tatatatgga
     2161 aaaggtcttg ttttgttaaa cttatgttag ttaactggat tcgtctttaa ccacaaaaag
     2221 ttttcaataa gctacaaatt tagacacgca agccgatgca gtcattagta catatattta
     2281 ttgcaagtga ttacatggca acccaaactt caaaaacagt aggttgctcc atttagtcgc
     2341 tgcaagaatt caagcttgga ggaaccgcaa cgttgaagga gccactcagc cgcgggtttc
     2401 tggagtttaa tgagctaagc acatacgtca gaaaccatta ttgcgcgttc aaaagtcgcc
     2461 taaggtcact atcagctagc aaatatttct tgtcaaaatg ctccactgac gttccataaa
     2521 ttcccctcgg tatccaatta gagtctcata ttcactctca atccaaataa tctgcaccgg
     2581 atctggatcg tttcgcatgt ctcctgaaag aaggcctgct gatattagaa gggctactga
     2641 ggctgatatg cctgctgtgt gcactattgt gaatcattac attgagactt ctactgtgaa
     2701 tttcaggact gagcctcaag aaccacaaga gtggactgat gatcttgtga ggcttagaga
     2761 gaggtatcct tggcttgttg ctgaagtgga tggtgaagtg gctggtattg cttatgctgg
     2821 tccttggaag gctaggaatg cttacgattg gactgctgag tctactgttt acgtgtctcc
     2881 taggcatcaa agaactggtc ttggatctac tctttacact catcttttga agtctttgga
     2941 ggctcagggt ttcaagtctg tggttgctgt gattggactt cctaatgatc cttctgtgag
     3001 gatgcatgag gctcttggtt atgctcctag gggaatgctt agagctgctg gtttcaaaca
     3061 tggaaattgg catgatgtgg gattctggca gcttgatttc tctcttcctg ttcctcctag
     3121 acctgttctt cctgtgactg agatttgagc gggactctgg atctagagtc aagcagatcg
     3181 ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat
     3241 tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac
     3301 gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat
     3361 agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt
     3421 actagatccg ctactagaat tcgagctcag cgttgtcaat caattggcaa gtcataaaat
     3481 gcattaaaaa atattttcat actcaactac aaatccatga gtataactat aattataaag
     3541 caatgattag aatctgacaa ggattctgga aaattacata aaggaaagtt cataaatgtc
     3601 taaaacacaa gaggacatac ttgtattcag taacatttgc agcttttcta ggtctgaaaa
     3661 tatatttgtt gcctagtgaa taagcataat ggtacaacta caagtgtttt actcctcata
     3721 ttaacttcgg tcattagagg ccacgatttg acacattttt actcaaaaca aaatgtttgc
     3781 atatctctta taatttcaaa ttcaacacac aacaaataag agaaaaaaca aataatatta
     3841 atttgagaat gaacaaaagg accatatcat tcattaactc ttctccatcc atttccattt
     3901 cacagttcga tagcgaaaac cgaataaaaa acacagtaaa ttacaagcac aacaaatggt
     3961 acaagaaaaa cagttttccc aatgccataa tactcaaact cagtaggatt ctggtgtgtg
     4021 cgcaatgaaa ctgatgcatt gaacttgacg aacgttgtcg aaaccgatga tacgaacgaa
     4081 agcaagcagc gttagccacc tccaaccttc ctcttcttct taggatcagc ccttgaatca
     4141 ccaccgagct gtgagagatc gatcctagtc tcgtagagtc cagtgataga ctgatggatg
     4201 agggtagcat cgagcacttc tttggtagac ctgtacacct tcctatcgat ggttgtatcg
     4261 aagtacttga aagccctagg agcaccgagg ttggtaaggg tgaagagatg gatgatgttc
     4321 tctgcctgtt ccctgatagg cttatctctg tgcttgttgt aagcagacaa caccttatcg
     4381 aggtttgcat cagcgaggat caccctttta gagaactcag agatctgctc gatgatctca
     4441 tccaagtagt gcttgtgctg ctcaacgaaa agttgcttct gctcgttatc ttctggagat
     4501 cccttcaact tctcgtagtg agaagcgagg taaagaaagt taacgtactt agatgggaga
     4561 gcaagctcgt ttcccttttg aaggaaccta gcagaagcga gcatcctctt tctaccgttc
     4621 tcgagttcga agagtgagta ctttgggagc ttgatgatga gatccttctt aacctctttg
     4681 tatcccttag cctcgaggaa atcgattggg ttcttctcga aagatgacct ttccatgata
     4741 gtgattccga gaagttcctt aacagacttg agcttcttac tctttccctt ctcaacctta
     4801 gccacaacga gaacagagta agccacggta ggagacacga aaccaccgta tttcttaggg
     4861 tcccaatcct tcttcctagc aatgagctta tcagagttcc tcttaggcct gatagactct
     4921 ttagagaatc caccggtctg cacctcggtt ttcttaacga tgttcacctg tggcatagag
     4981 agcacctttc taacggtagc gaaatccctt cccttatccc acacgatctc acctgtttca
     5041 ccgtttgtct cgatgagtgg cctctttctg atctcaccgt tagcgagggt aatctcggtc
     5101 ttgaagaaat tcatgatgtt agagtagaag aaatacttag cggtagcctt tccgatctct
     5161 tgctcagact tagcgatcat cttcctcaca tcgtacacct tgtaatcacc gtacacgaac
     5221 tctgactcga gcttaggata cttcttgatg agagcggttc caacaacagc gttaaggtaa
     5281 gcatcgtgag cgtggtggta gttgttgatt tccctcacct tgtagaattg gaaatccttt
     5341 ctgaaatcag acacgagctt tgacttgagg gtgataacct tcacttccct gatcaactta
     5401 tcgttctcat cgtacttggt gttcatccta gaatcgagga tctgtgcaac gtgcttagtg
     5461 atctgcctgg tttccacaag ctgcctcttg atgaatcctg ccttatccaa ttcagagagt
     5521 cctcccctct cagccttagt caagttatcg aactttctct gagtgatgag cttagcgttg
     5581 aggagctgcc tccaatagtt cttcattttc ttcacaacct cttcacttgg cacgttatca
     5641 ctcttacccc tgttcttatc agacctggtg agcaccttgt tatcgataga atcatccttc
     5701 aagaatgact gtggcacgat atgatcaaca tcgtaatcag agagcctgtt gatatccaac
     5761 tcttgatcca catacatatc ccttccgttc tggaggtagt agaggtagag cttctcattc
     5821 tggagctgag tgttctcaac agggtgctct ttgaggatct gagatccaag ctctttgata
     5881 ccttcctcga tcctcttcat cctttcccta gagttcttct gtcccttctg agtggtctgg
     5941 ttctctctag ccatttcgat cacgatgttc tcaggcttat gccttcccat caccttcacc
     6001 aactcatcca caaccttcac agtctggagg attcccttct tgattgcagg agatccagcg
     6061 aggttagcga tatgctcatg gagactatca ccctgtcctg aaacctgagc cttctggata
     6121 tcctctttaa aggtgagaga atcatcgtgg atgagctgca tgaagtttct gttagcgaat
     6181 ccatcagact tgaggaaatc aaggattgtc tttccagact gcttatccct gattccgtta
     6241 atgagctttc ttgagagcct tccccaacca gtgtatcttc ttctcttcaa ctgcttcatc
     6301 accttatcat cgaagagatg agcgtaggtc ttgagccttt cttcaatcat ctctctatct
     6361 tcaaagaggg tgagggtaag aacgatatcc tccaagatat cctcgttttc ctcgttatcc
     6421 aagaaatcct tatccttaat gatcttgagg agatcgtggt aggttccgag agatgcgttg
     6481 aacctatcct caacaccaga aatctcaact gaatcgaagc actcgatttt cttgaagtaa
     6541 tcctctttga gctgcttcac ggtcaccttt ctgttggtct tgaacaagag atcaacgata
     6601 gccttctttt gctcacctga caaaaaagca ggcttcctca ttccctcggt cacgtactta
     6661 accttggtca actcgttgta cacggtgaag tactcgtaga gcaaagagtg cttagggagc
     6721 accttctcgt ttggaaggtt cttatcgaag ttggtcatcc tctcgatgaa agactgagca
     6781 ctagcaccct tatccaccac ctcttcgaag ttccaagggg tgatggtttc ctcagacttt
     6841 ctggtcatcc aagcgaatct tgagtttcct ctagcgagag gtcccacgta gtaagggatt
     6901 ctgaaggtga gaatcttctc aatcttttcc ctgttatcct tgaggaatgg gtagaaatcc
     6961 tcttgccttc taaggatagc gtgcaactct ccgaggtgga tctgatgagg gatagatccg
     7021 ttatcgaagg tcctctgctt tctgagaaga tcctctctat tgagcttcac gaggagttcc
     7081 tcggttccat ccatcttctc gaggataggc ttgatgaact tgtagaactc ttcttgagat
     7141 gcaccaccat cgatgtaacc agcgtatccg ttcttagact gatcgaagaa aatctctttg
     7201 tacttctctg ggagctgctg tctaacaaga gccttgagaa gtgtgagatc ctggtggtgc
     7261 tcatcgtatc tcttgatcat agaagctgag agtggagcct tggtgatctc ggtgttcact
     7321 ctgaggatat cactgaggag gatagcatca gagaggttct tagcagcgag gaacaaatca
     7381 gcgtactgat ctccgatctg agcgaggagg ttatcgagat catcatcgta ggtatccttt
     7441 gagagctgga gctttgcatc ctcagcgaga tcgaagttag acttgaagtt aggggtgagt
     7501 ccgagagaga gagcgatcaa gtttccgaaa agtccgttct tcttctcacc agggagctga
     7561 gcaatgaggt tctcaagcct tcttgactta gagagcctag cagagaggat agccttagca
     7621 tccacacctg aagcgttgat agggttctct tcgaaaagct ggttgtaggt ctgcacgagc
     7681 tggatgaaca acttatccac atcagagtta tcagggttga gatcaccctc gatgaggaag
     7741 tgtcctctga acttgatcat gtgagcgaga gcgaggtaga tgagcctgag atcagcctta
     7801 tcagtagaat caacgagctt ctttctgagg tggtagatag tagggtactt ctcgtggtat
     7861 gccacctcat caacgatgtt tccgaagata gggtgcctct cgtgcttctt atcttcttcc
     7921 acgaggaatg actcttcgag cctgtggaag aatgaatcat ccactttagc catctcgtta
     7981 gagaagatct cttggaggta gcagatcctg ttctttcttc tggtgtacct tcttctagcg
     8041 gttctcttga gtctggtagc ctcagcagtt tcaccagaat cgaagaggag agcaccgata
     8101 aggtttttct tgatagagtg cctatcggtg tttccgagaa ccttgaactt cttagatggc
     8161 accttgtact catcggtgat cacagcccat cccacagagt tagttccgat ggcgagtccg
     8221 atagagtact tcttatccat tctgccgcca gagctgccac ccgacgattc tggcgtcgcg
     8281 ctctccgagg ttcccggtgt ctccgaacca gaggatcccc ctgacgagcc tccagaatcc
     8341 gtggaggact gggccttctt ctgagcattg aacacctgcc tcggcatgcg aaaaaagtag
     8401 caaagcagag cggcacactc gtccgcaaga atgccttctg ttatctcaac tctgtgattc
     8461 atcccggggt aatgaagcac gtccatcaga gatcctgccg ccccggtttt ggcgttgcga
     8521 actccaaaaa ccacgcgtcc gatccggcta tgaatcatag caccagcaca cataacgcaa
     8581 ggctcgaacg tgacatagag tgttgcgtca ataagcctat agttctgcat aaccagccct
     8641 ccctgcctca aagccataat ctctgcgtgc gccgtagggt cgtgcaatcc gattgcccta
     8701 ttccagcctt cgccaatcac cctattgttc aacaccagga ctgcgccaac aggaacctcg
     8761 cgttcgtctc tggcccgctt agccagagtc agggcgtgtc tcatccagta ctcatgggag
     8821 aattcaacct cggaagatcc gccgctagac ccaccagacg actctggtgt cgccgactcc
     8881 gacgtgcctg gagtctcaga acctgacgac cccccggagc tgccaccaga atccgtcgat
     8941 gactgtgcct tcttctgcgc tttgatttct tgtctccgca tccggaagaa gtcggaaagc
     9001 agagccgcgc attcatccgc caaaataccc tcggtgattt ccaccctgtg gttcatgcca
     9061 ggatggtgca aaacgtccat aaggctacct gctgcgcctg tttttgcatc cctagcgccg
     9121 aacacgacgc gcccaatgcg ggaatgaatc attgcgcccg cgcacatgac acacggctcg
     9181 agagtcacat aaagagtagc gtctatcaag cgatagttct gcatcaccag acccccctgt
     9241 ctaagagcca taatctcagc atgtgctgtt ggatcgtgtc tgcctatcgg ccgattccac
     9301 ccctctccga taacgcggtt attgtgcaca agcacagccc ccaccggaac ttcccgctcg
     9361 tcccacgctc tcttggccaa cgtcaaagcg tgccgcatcc aatactcgtg ggaaaactca
     9421 acttctgaca ttggctgtgg tgagagaaac agagcgtgag ctcaaataca caagtacgaa
     9481 atcagagtta tgagagagga tttggagaaa gaagatcacc tgaagatcgt ttgcgtcgcg
     9541 cgaatgagag agacggcaga gaaaacgaga agctttaaga ggacgaacaa aaaccctaat
     9601 gtcactcctt ttatatagat gagaaattta gggtttgggc tctaatggag ctgtctacat
     9661 gggctttcat ttctttatcc agtaaagaac cggtttaaca ggttccctgg tttatacgtg
     9721 gttttagccg gttaacttac ggaatattat ttttatctta agttacacaa attacatcat
     9781 taatagtaaa acaaaatttt atggtgcaat ttaagttttt atgatgagtg aaattcgaat
     9841 aatatacaaa tattttactg tttaggtttc cctaaaccta cttttagata tgtaaattaa
     9901 acattttttt ttttttggtt agaaccaagt cagtgatcat agaactcaaa aagctggtgt
     9961 ttcaattggt acaagcattt ttattgcaag atataaaaat tcagaaggac atatccttga
    10021 tgtatgtaca tgaaggaacc gattttgact ccaagctaca aagaagaaga agaatgatat
    10081 acaaagaaaa ctgcatatcc tcaccatgaa attgaaagct agaggaaggg gaataacaga
    10141 tactctgcgt agaagagatt ccatataaac cgataatatc ctgagatagc ttcctgcaat
    10201 aatcaaatgt ttctacactc cgttagatat acactccatt gatgttccgt gataacgagt
    10261 ttatgtttgt acaaaagtat tttaccttgg tgtagtttgc tttttctagt acccatgtct
    10321 accaaagatc aacaacaaag atattagaga ttaatgaatt tggcagtgtc aaatgcagaa
    10381 actaaagcat atagttatga agttcttgtg cattatctca aaagtatgta agagatataa
    10441 gagtatctca taatgcttta cctggaaaat taagcctgaa gccaagatca catgtgcagg
    10501 aatcatcaag ctacctctaa aaacctgttc attgttaaag gataagaggg ttggtctttt
    10561 caaagctgca tactgaggaa agtaacaata tcgcaacaaa aataaattgg tggcgatagt
    10621 ttatcaagct tctacatgca gtaatggtgt tcgtgcttta attaaccgaa atagttaaga
    10681 cggaaataga tgaattcacc tgaggcatgt agaaagctag tgatatggct gaaacataat
    10741 ttactagcag aagtccagaa ccgaggaatg caatgtttct cactccaagt tttgttgcca
    10801 gggttgatat ttggaaccta ttcaagatca atcaaaggca tgattaacta aacatgcttg
    10861 aacaaacaca taaagacaat atatcaaacg gagctggtta aacattgtct ttgactgtca
    10921 aatggattga gccaaagagc ggaaagaaaa tggttatgcg tgcagttact tactttcgat
    10981 ctccttcaac atcaggaagg tcctttgtaa tagcaatgac cagtgcaaac agtgtcacaa
    11041 aagatgtgat gaacgccaca ggtgcactgc aaatagcgaa tcaaaagttg agtaatatat
    11101 ctactcacga ttatggggag ctccttacga attcccatgg ggagcatctt cattcttaag
    11161 atatgaagat aatcttcaaa aggcccctgg gaatctgaaa gaagagaagc aggcccattt
    11221 atatgggaaa gaacaatagt atttcttata taggcccatt taagttgaaa acaatcttca
    11281 aaagtcccac atcgcttaga taagaaaacg aagctgagtt tatatacagc tagagtcgaa
    11341 gtagtgattg tgagaccgtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
    11401 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
    11461 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
    11521 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
    11581 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
    11641 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
    11701 tcgagctcgg tacccgggga tcctctagag tcgacctgca ggcatgcaag cttggcgtaa
    11761 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
    11821 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
    11881 attgcgttgc gctcactgcc cgctttccag tcggcaaacc tgtcgtgcca gctgcggtct
    11941 cagtttaaga gctatgctgg aaacagcata gcaagtttaa ataaggctag tccgttatca
    12001 acttgaaaaa gtggcaccga gtcggtgctt ttttttgcaa aattttccag atcgatttct
    12061 tcttcctctg ttcttcggcg ttcaatttct ggggtcgctc agagaggatg cacatgtgac
    12121 cgagggacac gaagtgatcc gtttaaacta tcagtgtttg acaggatata ttggcgggta
    12181 aacctaagag aaaagagcgt ttattagaat aatcggatat ttaaaagggc gtgaaaaggt
    12241 ttatccgttc gtccatttgt atgtgccagc cgtgcggctg catgaaatcc tggccggttt
    12301 gtctgatgcc aagctggcgg cctggccggc cagcttggcc gctgaagaaa ccgagcgccg
    12361 ccgtctaaaa aggtgatgtg tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta
    12421 tatgatgcga tgagtaaata aacaaatacg caaggggaac gcatgaaggt tatcgctgta
    12481 cttaaccaga aaggcgggtc aggcaagacg accatcgcaa cccatctagc ccgcgccctg
    12541 caactcgccg gggccgatgt tctgttagtc gattccgatc cccagggcag tgcccgcgat
    12601 tgggcggccg tgcgggaaga tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt
    12661 gaccgcgacg tgaaggccat cggccggcgc gacttcgtag tgatcgacgg agcgccccag
    12721 gcggcggact tggctgtgtc cgcgatcaag gcagccgact tcgtgctgat tccggtgcag
    12781 ccaagccctt acgacatatg ggccaccgcc gacctggtgg agctggttaa gcagcgcatt
    12841 gaggtcacgg atggaaggct acaagcggcc tttgtcgtgt cgcgggcgat caaaggcacg
    12901 cgcatcggcg gtgaggttgc cgaggcgctg gccgggtacg agctgcccat tcttgagtcc
    12961 cgtatcacgc agcgcgtgag ctacccaggc actgccgccg ccggcacaac cgttcttgaa
    13021 tcagaacccg agggcgacgc tgcccgcgag gtccaggcgc tggccgctga aattaaatca
    13081 aaactcattt gagttaatga ggtaaagaga aaatgagcaa aagcacaaac acgctaagtg
    13141 ccggccgtcc gagcgcacgc agcagcaagg ctgcaacgtt ggccagcctg gcagacacgc
    13201 cagccatgaa gcgggtcaac tttcagttgc cggcggagga tcacaccaag ctgaagatgt
    13261 acgcggtacg ccaaggcaag accattaccg agctgctatc tgaatacatc gcgcagctac
    13321 cagagtaaat gagcaaatga ataaatgagt agatgaattt tagcggctaa aggaggcggc
    13381 atggaaaatc aagaacaacc aggcaccgac gccgtggaat gccccatgtg tggaggaacg
    13441 ggcggttggc caggcgtaag cggctgggtt gtctgccggc cctgcaatgg cactggaacc
    13501 cccaagcccg aggaatcggc gtgagcggtc gcaaaccatc cggcccggta caaatcggcg
    13561 cggcgctggg tgatgacctg gtggagaagt tgaaggccgc gcaggccgcc cagcggcaac
    13621 gcatcgaggc agaagcacgc cccggtgaat cgtggcaagc ggccgctgat cgaatccgca
    13681 aagaatcccg gcaaccgccg gcagccggtg cgtcgtcgat taggaagccg cccaagggcg
    13741 acgagcaacc agattttttc gttccgatgc tctatgacgt gggcacccgc gatagtcgca
    13801 gcatcatgga cgtggccgtt ttccgtctgt cgaagcgtga ccgacgagct ggcgaggtga
    13861 tccgctacga gcttccagac gggcacgtag aggtttccgc agggccggcc ggcatggcca
    13921 gtgtgtggga ttacgacctg gtactgatgg cggtttccca tctaaccgaa tccatgaacc
    13981 gataccggga agggaaggga gacaagcccg gccgcgtgtt ccgtccacac gttgcggacg
    14041 tactcaagtt ctgccggcga gccgatggcg gaaagcagaa agacgacctg gtagaaacct
    14101 gcattcggtt aaacaccacg cacgttgcca tgcagcgtac gaagaaggcc aagaacggcc
    14161 gcctggtgac ggtatccgag ggtgaagcct tgattagccg ctacaagatc gtaaagagcg
    14221 aaaccgggcg gccggagtac atcgagatcg agctagctga ttggatgtac cgcgagatca
    14281 cagaaggcaa gaacccggac gtgctgacgg ttcaccccga ttactttttg atcgatcccg
    14341 gcatcggccg ttttctctac cgcctggcac gccgcgccgc aggcaaggca gaagccagat
    14401 ggttgttcaa gacgatctac gaacgcagtg gcagcgccgg agagttcaag aagttctgtt
    14461 tcaccgtgcg caagctgatc gggtcaaatg acctgccgga gtacgatttg aaggaggagg
    14521 cggggcaggc tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat
    14581 ccgccggttc ctaatgtacg gagcagatgc tagggcaaat tgccctagca ggggaaaaag
    14641 gtcgaaaaag cttctttcct gtggatagca cgtacattgg gaacccaaag ccgtacattg
    14701 ggaaccggaa cccgtacatt gggaacccaa agccgtacat tgggaaccgg tcacacatgt
    14761 aagtgactga tataaaagag aaaaaaggcg atttttccgc ctaaaactct ttaaaactta
    14821 ttaaaactct taaaacccgc ctggcctgtg cataactgtc tggccagcgc acagccgaac
    14881 agctgcaaaa agcgcccaca tcaaggctcc gagtgcgcgg aacccctatt tgtttatttt
    14941 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
    15001 aatattgaaa aaggaagagt atggctaaaa tgagaatatc accggaattg aaaaaactga
    15061 tcgaaaaata ccgctgcgta aaagatacgg aaggaatgtc tcctgctaag gtatataagc
    15121 tggtgggaga aaatgaaaac ctatatttaa aaatgacgga cagccggtat aaagggacca
    15181 cctatgatgt ggaacgggaa aaggacatga tgctatggct ggaaggaaag ctgcctgttc
    15241 caaaggtcct gcactttgaa cggcatgatg gctggagcaa tctgctcatg agtgaggccg
    15301 atggcgtcct ttgctcggaa gagtatgaag atgaacaaag ccctgaaaag attatcgagc
    15361 tgtatgcgga gtgcatcagg ctctttcact ccatcgacat atcggattgt ccctatacga
    15421 atagcttaga cagccgctta gccgaattgg attacttact gaataacgat ctggccgatg
    15481 tggattgcga aaactgggaa gaggacactc catttaaaga tccgcgcgag ctgtatgatt
    15541 ttttaaagac ggaaaagccc gaagaggaac ttgtcttttc ccacggcgac ctgggagaca
    15601 gcaacatctt tgtgaaagat ggcaaagtaa gtggctttat tgatcttggg agaagcggca
    15661 gggcggacaa gtggtatgac attgccttct gcgtccggtc gctcagggag gatatcgggg
    15721 aagaacagta tgtcgagcta ttttttgact tactggggat caagcctgat tgggagaaaa
    15781 taaaatatta tattttactg gatgaattgt tttagctgtc agaccaagtt tactcatata
    15841 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
    15901 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
    15961 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
    16021 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
    16081 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
    16141 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
    16201 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
    16261 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
    16321 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
    16381 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
    16441 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
    16501 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
    16561 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctgctc ggatctgttg
    16621 gaccggacag tagtcatggt tgatgggctg cctgtatcga gtggtgattt tgtgccgagc
    16681 tgccggtcgg ggagctgttg gctggctggt ggcaggatat attgtggtgt aaacaaattg
    16741 acgcttagac aacttaataa cacattgcgg acgtttttaa tgtactgggg ttgaacactc
    16801 tg
//
