LOCUS       Exported               12704 bp ds-DNA     circular SYN 02-JUN-2022
DEFINITION  for CRISPRi, lentiviral expression of KRAB-dCas9 and BlastR. Also 
            called pXPR_121.
ACCESSION   .
VERSION     .
KEYWORDS    pLX_311-KRAB-dCas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12704)
  AUTHORS   Rosenbluh J, Xu H, Harrington W, Gill S, Wang X, Vazquez F, Root DE,
            Tsherniak A, Hahn WC
  TITLE     Complementary information derived from CRISPR Cas9 mediated gene 
            deletion and suppression.
  JOURNAL   Nat Commun. 2017 May 23;8:15403. doi: 10.1038/ncomms15403.
  PUBMED    28534478
REFERENCE   2  (bases 1 to 12704)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 2, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12704
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        98..1276
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          329..1267
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1224..1244
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1296..1320
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1350..1535
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             1557..5660
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5664..5690
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5709..5729
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5736..5756
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(5801..5825)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5833..5874
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    5925..6513
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5978..5998)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6396..6407)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6585..6818
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6896..7017
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6933..6952)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6987..7006
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7057..7192
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7212..7231)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7213..7231)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7241..7258)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7241..7257)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7250..7272)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7399..7854
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7486..7505)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7696..7717
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7880..7984
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7985..8845
                     /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(8203..8222)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9016..9604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9505..9524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9758..9775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9892..9913
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9928..9958
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9966..9982
                     /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)."
     primer_bind     9971..9993
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9990..10006
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9990..10006
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10025..10045
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10027..10045
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        10073..10299
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             10300..10480
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10527..10652
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11145..11378
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11563..11607
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        11838..12048
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        11860..12056
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      11907..12042
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             12078..12473
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    12547..12664
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggctcgaggc ctgcaggtgc aaagatggat aaagttttaa acagagagga atctttgcag
       61 ctaatggacc ttctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc
      121 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg
      181 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
      241 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
      301 tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg
      361 cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc tgcagtacgt
      421 gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc cttgcgctta
      481 aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg gccgccgcgt
      541 gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta
      601 aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg
      661 ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc
      721 gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg
      781 ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc
      841 cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc
      901 gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg gagagcgggc
      961 gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga
     1021 ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct tttggagtac
     1081 gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt
     1141 ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt
     1201 tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc
     1261 atttcaggtg tcgtgaggct agcatcgatt gatcaacaag tttgtacaaa aaagttggca
     1321 tggatgctaa gtcactaact gcctggtccc ggacactggt gaccttcaag gatgtatttg
     1381 tggacttcac cagggaggag tggaagctgc tggacactgc tcagcagatc gtgtacagaa
     1441 atgtgatgct ggagaactat aagaacctgg tttccttggg ttatcagctt actaagccag
     1501 atgtgatcct ccggttggag aagggagaag agcccggcgg ttccggcgga gggtcgatgg
     1561 acaagaagta cagcatcggc ctggccatcg gcaccaactc tgtgggctgg gccgtgatca
     1621 ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca
     1681 gcatcaagaa gaacctgatc ggcgccctgc tgttcgacag cggagaaaca gccgaggcca
     1741 cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc
     1801 tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg
     1861 aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca
     1921 tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac
     1981 tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga
     2041 tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg
     2101 acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca
     2161 acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc
     2221 tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggcaacctga
     2281 ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg
     2341 ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga
     2401 tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc
     2461 tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga
     2521 tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc
     2581 agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct
     2641 acatcgatgg cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa
     2701 agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc
     2761 agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca
     2821 ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga
     2881 agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat
     2941 tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg
     3001 tggacaaggg cgccagcgcc cagagcttca tcgagcggat gaccaacttc gataagaacc
     3061 tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtaca
     3121 acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg
     3181 gcgagcagaa aaaagccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga
     3241 agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg
     3301 gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca
     3361 aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga
     3421 ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc
     3481 tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc
     3541 tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt
     3601 tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc
     3661 tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg
     3721 agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga
     3781 aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg
     3841 aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga
     3901 agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg
     3961 aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata
     4021 tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gacgctatcg
     4081 tgcctcagag ctttctgaag gacgactcca tcgataacaa agtgctgact cggagcgaca
     4141 agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact
     4201 actggcgcca gctgctgaat gccaagctga ttacccagag gaagttcgac aatctgacca
     4261 aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg
     4321 tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta
     4381 agtacgacga gaacgacaaa ctgatccggg aagtgaaagt gatcaccctg aagtccaagc
     4441 tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc
     4501 accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc
     4561 ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga
     4621 tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca
     4681 tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc
     4741 tgatcgagac aaacggcgaa acaggcgaga tcgtgtggga taagggccgg gactttgcca
     4801 ccgtgcggaa agtgctgtct atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga
     4861 caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgacaag ctgatcgcca
     4921 gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt
     4981 ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag
     5041 agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc
     5101 tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact
     5161 ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga
     5221 agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact
     5281 atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc
     5341 acaaacacta cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc
     5401 tggccgacgc taatctggac aaggtgctga gcgcctacaa caagcacaga gacaagccta
     5461 tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg
     5521 ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg
     5581 tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc
     5641 tgtctcagct gggaggcgac gcctatccct atgacgtgcc cgattatgcc agcctgggca
     5701 gcggctcccc caagaaaaaa cgcaaggtgg aagatcctaa gaaaaagcgg aaagtggacg
     5761 gcattggtag tgggagcaac ggcagcagcg gatcctaaac ccccaacttt cttgtacaaa
     5821 gtggttgata tcggtaagcc tatccctaac cctctcctcg gtctcgattc tacgtagtaa
     5881 tgaactagta ccggttaagt cgacaatcaa cgcgttaagt cgacaatcaa cctctggatt
     5941 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     6001 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     6061 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     6121 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     6181 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     6241 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     6301 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     6361 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     6421 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     6481 cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa
     6541 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     6601 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     6661 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     6721 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     6781 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt acgtatagta gttcatgtca
     6841 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     6901 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6961 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     7021 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     7081 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     7141 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg
     7201 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     7261 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     7321 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     7381 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     7441 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     7501 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     7561 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     7621 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     7681 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     7741 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     7801 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     7861 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     7921 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     7981 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     8041 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     8101 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     8161 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     8221 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     8281 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     8341 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     8401 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     8461 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     8521 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     8581 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     8641 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     8701 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     8761 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     8821 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     8881 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     8941 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     9001 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     9061 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     9121 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     9181 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     9241 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     9301 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     9361 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     9421 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     9481 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     9541 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     9601 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     9661 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     9721 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     9781 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     9841 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     9901 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     9961 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
    10021 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct
    10081 tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa
    10141 ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta
    10201 ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc
    10261 agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag
    10321 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
    10381 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
    10441 tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact
    10501 tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc
    10561 gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag
    10621 gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc
    10681 gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag
    10741 tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag
    10801 aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac
    10861 ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa
    10921 aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg
    10981 cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga
    11041 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag
    11101 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt
    11161 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag
    11221 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag
    11281 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc
    11341 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga
    11401 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa
    11461 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc
    11521 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta
    11581 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg
    11641 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct
    11701 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac
    11761 ctcccaaccc cgaggggacc catgcattgc atctcaatta gtcagcaacc aggtgtggaa
    11821 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcgtctcaat tagtcagcaa
    11881 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
    11941 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
    12001 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
    12061 tttctagagg taccaccatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa
    12121 gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag
    12181 ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac
    12241 cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt
    12301 gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac
    12361 aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc
    12421 cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggcctgcagc
    12481 tgcagtagta aggcgcgccg ttaacgaatt ctagatcttg agacaaatgg cagtattcat
    12541 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
    12601 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
    12661 ttttcgggtt tattacaggg acagcagaga tccactttgg cgcc
//
