LOCUS       sequence-199012-       15807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for CRISPRi, lentiviral expression of dCas9-p300 2A BlastR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15807)
  AUTHORS   Najm FJ, Strand C, Donovan KF, Hegde M, Sanson KR, Vaimberg EW, 
            Sullender ME, Hartenian E, Kalani Z, Fusi N, Listgarten J, Younger 
            ST, Bernstein BE, Root DE, Doench JG
  TITLE     Orthologous CRISPR-Cas9 enzymes for combinatorial genetic screens.
  JOURNAL   Nat Biotechnol. 2018 Feb;36(2):179-189. doi: 10.1038/nbt.4048. Epub 
            2017 Dec 18.
  PUBMED    29251726
REFERENCE   2  (bases 1 to 15807)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nbt.4048"; 
            journalName: "Nat Biotechnol"; date: "2018-02"; volume: "36"; issue:
            "2"; pages: "179-189"
COMMENT     Sequence Label: pXPR_123
FEATURES             Location/Qualifiers
     source          1..15807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      248..676
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(335..354)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     545..566
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(685..705)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        690..1019
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      870..1005
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     932..951
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1067..1114
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1133..1507
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    1637..1758
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1674..1693)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1728..1747
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1807..1823)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1807..1823)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1820..1842)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1831..1847
                     /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)."
     promoter        complement(join(1855..1861,1862..1879,1880..1885))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1900..1921
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2038..2055)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2209..2797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2289..2308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2968..3759,3760..3828))
                     /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     3591..3610
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3829..3933)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4008..4027)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4199..4578
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4580..4778
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4733..4753
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4796..4976
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5023..5148
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5641..5874
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6059..6103
                     /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"
     misc_feature    6401..6518
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6648..7826
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6879..7817
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7774..7794
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             7842..7889
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(7890..7913,7914..7916,7917..10472,10473..10475,
                     10476..11990)
                     /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 N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             12045..12065
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12081..13931
                     /codon_start=1
                     /gene="EP300"
                     /product="catalytic histone acetyltransferase core of the 
                     human E1A-associated protein p300 (Delvecchio et al., 
                     2013)"
                     /label=p300 core
                     /translation="IFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDI
                     VKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQ
                     EIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQG
                     ESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAG
                     FVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHA
                     SDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPP
                     PNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDY
                     IFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFE
                     GDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLS
                     RGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDL
                     MDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQD"
     regulatory      13695..13704
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             13932..13958
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             13983..14036
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             14037..14435
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    14460..15048
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(14513..14533)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(14931..14942)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(15051..15067)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(15052..15068)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             15573..15753
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(15779..15796)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    join(15785..15807,1..202)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
       61 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
      121 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
      181 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
      241 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
      301 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
      361 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
      421 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
      481 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
      541 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
      601 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
      661 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      721 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      781 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      841 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      901 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
      961 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     1021 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc
     1081 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa
     1141 gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg
     1201 gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg
     1261 ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct
     1321 ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc
     1381 cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg
     1441 gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca
     1501 ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     1561 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     1621 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     1681 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1741 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     1801 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     1861 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     1921 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1981 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     2041 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2101 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2161 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2221 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2281 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2341 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2401 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2461 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2521 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2581 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2641 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2701 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2761 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2821 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2881 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2941 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3001 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3061 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3121 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3181 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3241 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3301 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3361 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3421 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3481 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3541 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3601 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3661 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3721 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3781 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3841 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3901 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     3961 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     4021 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     4081 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     4141 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
     4201 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     4261 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     4321 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     4381 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     4441 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     4501 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     4561 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     4621 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     4681 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     4741 ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc
     4801 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     4861 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     4921 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
     4981 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
     5041 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     5101 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     5161 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     5221 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     5281 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     5341 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     5401 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     5461 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
     5521 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     5581 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     5641 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     5701 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     5761 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     5821 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     5881 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     5941 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     6001 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     6061 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     6121 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     6181 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     6241 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     6301 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg
     6361 tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     6421 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     6481 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     6541 gagatccagt ttggttaatt agctagctgc aaagatggat aaagttttaa acagagagga
     6601 atctttgcag ctaatggacc ttctaggtct tgaaaggagt gggaattggc tccggtgccc
     6661 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     6721 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     6781 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     6841 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
     6901 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc
     6961 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
     7021 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
     7081 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
     7141 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
     7201 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
     7261 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
     7321 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
     7381 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
     7441 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
     7501 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
     7561 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     7621 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     7681 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     7741 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     7801 tttttcttcc atttcaggtg tcgtgacgta cggccaccat gaaaaggccg gcggccacga
     7861 aaaaggccgg ccaggcaaaa aagaaaaagg acaagaagta cagcatcggc ctggccatcg
     7921 gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat
     7981 tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc
     8041 tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat
     8101 acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca
     8161 aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga
     8221 agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt
     8281 accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc
     8341 ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg
     8401 gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct
     8461 acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc
     8521 tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg
     8581 agaagaagaa tggcctgttc ggcaacctga ttgccctgag cctgggcctg acccccaact
     8641 tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg
     8701 acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg
     8761 ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga
     8821 tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc
     8881 tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct
     8941 tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt
     9001 tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga
     9061 agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc
     9121 accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat
     9181 tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg
     9241 tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa
     9301 ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca
     9361 tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca
     9421 gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg
     9481 agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc
     9541 tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa
     9601 tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg
     9661 gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa
     9721 acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga
     9781 tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga
     9841 agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg
     9901 acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa
     9961 acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc
    10021 aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg
    10081 ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg
    10141 gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga
    10201 agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg
    10261 gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt
    10321 acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc
    10381 ggctgtccga ctacgatgtg gaccacatcg tgcctcagag ctttctgaag gacgactcca
    10441 tcgacaacaa ggtgctgacc agaagcgaca aggcccgggg caagagcgac aacgtgccct
    10501 ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga
    10561 ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg
    10621 ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg
    10681 cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg
    10741 aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt
    10801 tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg
    10861 tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg
    10921 actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg
    10981 ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc
    11041 tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga
    11101 tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag
    11161 tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc
    11221 ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg
    11281 gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg
    11341 gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa
    11401 gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa
    11461 aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga
    11521 gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat
    11581 atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata
    11641 atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc
    11701 agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt
    11761 ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc
    11821 tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg
    11881 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca
    11941 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac agcgctggag
    12001 gaggtggaag cggaggagga ggaagcggag gaggaggtag cggacctaag aaaaagagga
    12061 aggtggcggc cgctggatcc attttcaaac cagaagaact acgacaggca ctgatgccaa
    12121 ctttggaggc actttaccgt caggacccag aatcccttcc ctttcgtcaa cctgtggacc
    12181 ctcagctttt aggaatccct gattactttg atattgtgaa gagccccatg gatctttcta
    12241 ccattaagag gaagttagac actggacagt atcaggagcc ctggcagtat gtcgatgata
    12301 tttggcttat gttcaataat gcctggttat ataaccggaa aacatcacgg gtatacaaat
    12361 actgctccaa gctctctgag gtctttgaac aagaaattga cccagtgatg caaagccttg
    12421 gatactgttg tggcagaaag ttggagttct ctccacagac actgtgttgc tacggcaaac
    12481 agttgtgcac aatacctcgt gatgccactt attacagtta ccagaacagg tatcatttct
    12541 gtgagaagtg tttcaatgag atccaagggg agagcgtttc tttgggggat gacccttccc
    12601 agcctcaaac tacaataaat aaagaacaat tttccaagag aaaaaatgac acactggacc
    12661 ctgaactgtt tgttgaatgt acggagtgcg gaagaaagat gcatcagatc tgtgtccttc
    12721 accatgagat catctggcct gctggattcg tctgtgatgg ctgtttaaag aaaagtgcac
    12781 gaactaggaa agaaaataag ttttctgcta aaaggttgcc atctaccaga cttggcacct
    12841 ttctagagaa tcgtgtgaat gactttctga ggcgacagaa tcaccctgag tcaggagagg
    12901 tcactgttag agtagttcat gcttctgaca aaaccgtgga agtaaaacca ggcatgaaag
    12961 caaggtttgt ggacagtgga gagatggcag aatcctttcc ataccgaacc aaagccctct
    13021 ttgcctttga agaaattgat ggtgttgacc tgtgcttctt tggcatgcat gttcaagagt
    13081 atggctctga ctgccctcca cccaaccaga ggagagtata catatcttac ctcgatagtg
    13141 ttcatttctt ccgtcctaaa tgcttgagga ctgcagtcta tcatgaaatc ctaattggat
    13201 atttagaata tgtcaagaaa ttaggttaca caacagggca tatttgggca tgtccaccaa
    13261 gtgagggaga tgattatatc ttccattgcc atcctcctga ccagaagata cccaagccca
    13321 agcgactgca ggaatggtac aaaaaaatgc ttgacaaggc tgtatcagag cgtattgtcc
    13381 atgactacaa ggatattttt aaacaagcta ctgaagatag attaacaagt gcaaaggaat
    13441 tgccttattt cgagggtgat ttctggccca atgttctgga agaaagcatt aaggaactgg
    13501 aacaggagga agaagagaga aaacgagagg aaaacaccag caatgaaagc acagatgtga
    13561 ccaagggaga cagcaaaaat gctaaaaaga agaataataa gaaaaccagc aaaaataaga
    13621 gcagcctgag taggggcaac aagaagaaac ccgggatgcc caatgtatct aacgacctct
    13681 cacagaaact atatgccacc atggagaagc ataaagaggt cttctttgtg atccgcctca
    13741 ttgctggccc tgctgccaac tccctgcctc ccattgttga tcctgatcct ctcatcccct
    13801 gcgatctgat ggatggtcgg gatgcgtttc tcacgctggc aagggacaag cacctggagt
    13861 tctcttcact ccgaagagcc cagtggtcca ccatgtgcat gctggtggag ctgcacacgc
    13921 agagccagga ctacccgtac gacgttccgg actacgcttc tgatatctgt acaggcagtg
    13981 gagagggcag aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct ggcccaatgg
    14041 ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca
    14101 gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca
    14161 tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc
    14221 tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg
    14281 agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc
    14341 ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg
    14401 aattgctgcc ctctggttat gtgtgggagg gctaagaatt cgatatcaag cttatcggta
    14461 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
    14521 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
    14581 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
    14641 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
    14701 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
    14761 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
    14821 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
    14881 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
    14941 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
    15001 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
    15061 gacctcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg
    15121 ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc
    15181 aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
    15241 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt
    15301 ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca gggccaggga
    15361 tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag caagagaagg
    15421 tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc ctgcatggga
    15481 tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta gcatttcatc
    15541 acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac cagatctgag
    15601 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
    15661 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
    15721 gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg ctgatcagcc
    15781 tcgactgtgc cttctagttg ccagcca
//
