LOCUS       sequence-431379-       13709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPRi, EF1a-driven dCas9-KRAB (ZNF10) (Cas only).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13709)
  AUTHORS   McGee AV, Liu YV, Griffith AL, Szegletes ZM, Wen B, Kraus C, Miller 
            NW, Steger RJ, Escude Velasco B, Bosch JA, Zirin JD, Viswanatha R, 
            Sontheimer EJ, Goodale A, Greene MA, Green TM, Doench JG
  TITLE     Modular vector assembly enables rapid assessment of emerging CRISPR 
            technologies.
  JOURNAL   Cell Genom. 2024 Mar 13;4(3):100519. doi: 
            10.1016/j.xgen.2024.100519.
  PUBMED    38484704
REFERENCE   2  (bases 1 to 13709)
  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"; journalName: "Cell 
            Genom."; date: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: pRDB_181
FEATURES             Location/Qualifiers
     source          1..13709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2640..3818
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2871..3809
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3766..3786
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3823..3832
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3868..3888
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3895..3918,3919..3921,3922..6477,6478..6480,
                     6481..7995)
                     /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             8026..8073
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8113..8307
                     /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
                     QLTKPDVILRLEKGEEPWLV"
     CDS             8332..8385
                     /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             8392..8790
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8801..9389
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8854..8874)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9272..9283)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9461..9694
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9720..9737)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9726..9950
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9996..10424
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10083..10102)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10293..10314
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10433..10453)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10438..10767
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10618..10753
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10680..10699
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10815..10862
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10881..11255
                     /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    11385..11506
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11422..11441)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11476..11495
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11555..11571)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11555..11571)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11568..11590)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11579..11595
                     /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(11603..11609,11610..11627,11628..11633))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11648..11669
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11786..11803)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11957..12545)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12037..12056)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12716..13507,13508..13576))
                     /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     13339..13358
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13577..13681)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta aggtaccggg cagaggtctc gcttttttcg agacaacacg
     2641 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     2701 gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     2761 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     2821 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
     2881 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
     2941 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg
     3001 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc
     3061 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt
     3121 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc
     3181 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg
     3241 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag
     3301 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg
     3361 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag
     3421 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga
     3481 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt
     3541 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt
     3601 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg
     3661 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat
     3721 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag
     3781 tggttcaaag tttttttctt ccatttcagg tgtcgtgaag gagccaccat ggccaaacgg
     3841 acagccgacg gaagcgagtt cgagtcacca aagaagaagc ggaaagtcgg cagcgacaag
     3901 aagtacagca tcggcctggc catcggcacc aactctgtgg gctgggccgt gatcaccgac
     3961 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
     4021 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
     4081 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
     4141 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
     4201 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
     4261 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
     4321 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
     4381 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     4441 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgct
     4501 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
     4561 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggcaa cctgattgcc
     4621 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
     4681 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     4741 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     4801 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     4861 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     4921 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
     4981 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     5041 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
     5101 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     5161 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     5221 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     5281 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     5341 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     5401 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
     5461 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
     5521 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     5581 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
     5641 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
     5701 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
     5761 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
     5821 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
     5881 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
     5941 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
     6001 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
     6061 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
     6121 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
     6181 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
     6241 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc agtggaaaac
     6301 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
     6361 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct
     6421 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaaggcc
     6481 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
     6541 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
     6601 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
     6661 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
     6721 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
     6781 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
     6841 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
     6901 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
     6961 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
     7021 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
     7081 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
     7141 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
     7201 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag
     7261 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
     7321 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
     7381 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
     7441 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
     7501 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
     7561 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
     7621 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
     7681 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
     7741 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
     7801 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
     7861 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
     7921 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacgaat cgatttgtct
     7981 cagctgggag gcgatgggtc cgggagcggc gggagcggag ggagcaaaag gccggcggcc
     8041 acgaaaaagg ccggccaggc aaaaaagaaa aagagcggag gctccgacgc caagagcctg
     8101 accgcctgga gcagaaccct ggtgaccttc aaggacgtgt tcgtggactt caccagagag
     8161 gagtggaagc tgctggacac cgcccagcag atcgtgtaca gaaacgtgat gctggagaac
     8221 tacaagaacc tggtgagcct gggctaccag ctgaccaagc ccgacgtgat cctgagactg
     8281 gagaagggcg aggagccctg gctggtggag agagagatcc accagggaag cgagggcaga
     8341 ggaagccttc taacatgcgg tgacgtggag gagaatcccg gcccttccgg gatggccaag
     8401 cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc
     8461 cccatctctg aggactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc
     8521 actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc
     8581 actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac
     8641 aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg
     8701 atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg
     8761 ctgccctctg gttatgtgtg ggagggctaa taaattcgaa aatcaacctc tggattacaa
     8821 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     8881 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     8941 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     9001 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     9061 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     9121 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     9181 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     9241 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     9301 ccgcggcctg ctgccggctc tgcggcctct tccgcgtcct cgccttcgcc ctcagacgag
     9361 tcggatctcc ctttgggccg cctccccgcg tcgactttaa gaccaatgac ttacaaggca
     9421 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     9481 caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     9541 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     9601 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     9661 agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttaaaccc gctgatcagc
     9721 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     9781 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     9841 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     9901 ggattgggaa gataatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     9961 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
    10021 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
    10081 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
    10141 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
    10201 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
    10261 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
    10321 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
    10381 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
    10441 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
    10501 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
    10561 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
    10621 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
    10681 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
    10741 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
    10801 tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg
    10861 acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc
    10921 gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg
    10981 tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc
    11041 aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc
    11101 tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca
    11161 tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca
    11221 actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca
    11281 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
    11341 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
    11401 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
    11461 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
    11521 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
    11581 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
    11641 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
    11701 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
    11761 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
    11821 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
    11881 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
    11941 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
    12001 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
    12061 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
    12121 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
    12181 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
    12241 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
    12301 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
    12361 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
    12421 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
    12481 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
    12541 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
    12601 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
    12661 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
    12721 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
    12781 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
    12841 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
    12901 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
    12961 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
    13021 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
    13081 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
    13141 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
    13201 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
    13261 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
    13321 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
    13381 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
    13441 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
    13501 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
    13561 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
    13621 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
    13681 gcacatttcc ccgaaaagtg ccacctgac
//
