LOCUS       sequence-431382-       13619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPRa, EF1a-driven dCas9-VP64 (Cas only).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13619)
  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 13619)
  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: pRDA_831
FEATURES             Location/Qualifiers
     source          1..13619
                     /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             8086..8235
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             8242..8295
                     /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             8302..8700
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8711..9299
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8764..8784)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9182..9193)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9371..9604
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9630..9647)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9636..9860
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9906..10334
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9993..10012)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10203..10224
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10343..10363)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10348..10677
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10528..10663
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10590..10609
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10725..10772
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10791..11165
                     /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    11295..11416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11332..11351)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11386..11405
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11465..11481)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11465..11481)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11478..11500)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11489..11505
                     /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(11513..11519,11520..11537,11538..11543))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11558..11579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11696..11713)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11867..12455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11947..11966)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12626..13417,13418..13486))
                     /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     13249..13268
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13487..13591)
                     /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 attggacgat
     8101 tttgatctgg atatgctggg aagtgacgcc ctcgatgatt ttgaccttga catgcttggt
     8161 tcggatgccc ttgatgactt tgacctcgac atgctcggca gtgacgccct tgatgatttc
     8221 gacctggaca tgctgggaag cgagggcaga ggaagccttc taacatgcgg tgacgtggag
     8281 gagaatcccg gcccttccgg gatggccaag cctttgtctc aagaagaatc caccctcatt
     8341 gaaagagcaa cggctacaat caacagcatc cccatctctg aggactacag cgtcgccagc
     8401 gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg
     8461 ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg
     8521 acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgc
     8581 cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga cagtgatgga
     8641 cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa
     8701 taaattcgaa aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     8761 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     8821 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     8881 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     8941 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     9001 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     9061 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     9121 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     9181 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     9241 tccgcgtcct cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg
     9301 tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     9361 aaggggggac tggaagggct aattcactcc caacgaagat aagatctgct ttttgcttgt
     9421 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     9481 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     9541 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     9601 agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     9661 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     9721 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     9781 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gataatagca ggcatgctgg
     9841 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     9901 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     9961 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
    10021 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
    10081 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
    10141 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
    10201 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
    10261 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
    10321 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
    10381 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
    10441 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
    10501 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
    10561 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
    10621 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
    10681 ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat
    10741 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt
    10801 tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga
    10861 ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg
    10921 acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg
    10981 cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca
    11041 cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc
    11101 gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg
    11161 actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
    11221 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
    11281 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
    11341 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
    11401 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
    11461 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
    11521 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
    11581 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
    11641 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
    11701 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
    11761 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
    11821 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
    11881 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
    11941 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
    12001 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
    12061 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
    12121 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
    12181 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
    12241 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
    12301 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
    12361 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
    12421 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
    12481 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
    12541 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
    12601 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
    12661 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
    12721 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
    12781 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
    12841 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
    12901 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
    12961 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
    13021 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
    13081 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
    13141 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
    13201 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
    13261 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
    13321 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
    13381 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
    13441 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
    13501 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
    13561 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgac
//
