LOCUS       sequence-231720-       13593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EF1a-ProCas9-Flavi-P2A-Puro.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13593)
  AUTHORS   Oakes BL, Fellmann C, Rishi H, Taylor KL, Ren SM, Nadler DC, Yokoo 
            R, Arkin AP, Doudna JA, Savage DF
  TITLE     CRISPR-Cas9 Circular Permutants as Programmable Scaffolds for Genome
            Modification.
  JOURNAL   Cell. 2019 Jan 10;176(1-2):254-267.e16. doi: 
            10.1016/j.cell.2018.11.052.
  PUBMED    30633905
REFERENCE   2  (bases 1 to 13593)
  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."; 
            date: "2019-01-10"; volume: "176(1-2):254-267.e16. doi"; pages: " 
            10.1016/j.cell.2018.11.052"
COMMENT     Sequence Label: pCF730
FEATURES             Location/Qualifiers
     source          1..13593
                     /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        2632..3809
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2863..3800
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3757..3777
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3816..3825
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3828..3848
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4866..7364
                     /codon_start=1
                     /product="C-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(C)
                     /translation="KPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEI
                     SGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTY
                     AHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH
                     DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE
                     NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL
                     QNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEV
                     VKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQI
                     LDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVV
                     GTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITL
                     ANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESIL
                     PKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIME
                     RSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALP
                     SKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLD
                     KVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATL
                     IHQSITGLYETRIDLSQLGGD"
     CDS             7986..8006
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8016..8072
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             8073..8669
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     8566..8586
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8685..9273
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8738..8758)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9156..9167)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9345..9578
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9604..9621)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9610..9834
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9880..10308
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9967..9986)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10177..10198
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10317..10337)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10322..10651
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10502..10637
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10564..10583
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10699..10746
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10765..11139
                     /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    11269..11390
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11306..11325)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11360..11379
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11439..11455)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11439..11455)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11452..11474)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11463..11479
                     /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(11487..11493,11494..11511,11512..11517))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11532..11553
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11670..11687)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11841..12429)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11921..11940)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12600..13391,13392..13460))
                     /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     13223..13242
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13461..13565)
                     /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 agctagctag gtcttgaaag gagtgggaat tggctccggt
     2641 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     2701 ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     2761 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     2821 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt
     2881 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccact
     2941 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
     3001 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
     3061 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
     3121 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
     3181 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
     3241 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
     3301 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
     3361 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     3421 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     3481 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     3541 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     3601 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     3661 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     3721 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     3781 gtttttttct tccatttcag gtgtcgtgaa ccggtgccac catggcccca aagaagaagc
     3841 ggaaggtcag cgctaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg
     3901 ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga
     3961 agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga
     4021 gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg
     4081 acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca
     4141 agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca
     4201 aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc
     4261 tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc
     4321 agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca
     4381 agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga
     4441 acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga
     4501 tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga
     4561 aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc
     4621 ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca
     4681 ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc
     4741 ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc
     4801 tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa
     4861 tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca
     4921 agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt
     4981 gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat
     5041 accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg
     5101 acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg
     5161 aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc
     5221 ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc
     5281 agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca
     5341 tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt
     5401 ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta
     5461 agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc
     5521 acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac
     5581 agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc
     5641 agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt
     5701 actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt
     5761 ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca
     5821 acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag
     5881 aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc
     5941 agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg
     6001 ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga
     6061 tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga
     6121 aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca
     6181 aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg
     6241 gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca
     6301 aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg
     6361 ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca
     6421 acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt
     6481 gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata
     6541 tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga
     6601 ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct
     6661 tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt
     6721 ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct
     6781 tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc
     6841 tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc
     6901 tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga
     6961 acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc
     7021 agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca
     7081 gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct
     7141 acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta
     7201 ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga
     7261 agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg
     7321 gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaaacaa aagaaacgcg
     7381 gaggcaagga caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg
     7441 ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg
     7501 accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag
     7561 ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga
     7621 tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc
     7681 acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct
     7741 tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga
     7801 gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg
     7861 cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca
     7921 gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa
     7981 ctagtccaaa aaagaagaga aaggtaggat ccggcgcaac aaacttctct ctgctgaaac
     8041 aagccggaga tgtcgaagag aatcctggac cgaccgagta caagcccacg gtgcgcctcg
     8101 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     8161 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     8221 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     8281 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     8341 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     8401 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggagtct
     8461 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     8521 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     8581 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     8641 ggtgcatgac ccgcaagccc ggtgcctgaa cgcgttaagt cgacaatcaa cctctggatt
     8701 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     8761 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     8821 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     8881 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     8941 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     9001 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     9061 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     9121 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     9181 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     9241 cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa
     9301 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     9361 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     9421 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     9481 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     9541 cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat
     9601 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     9661 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     9721 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     9781 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     9841 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     9901 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     9961 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
    10021 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
    10081 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
    10141 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
    10201 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
    10261 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
    10321 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
    10381 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
    10441 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
    10501 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
    10561 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
    10621 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
    10681 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
    10741 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
    10801 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
    10861 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
    10921 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
    10981 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
    11041 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
    11101 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
    11161 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
    11221 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
    11281 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
    11341 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
    11401 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
    11461 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
    11521 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
    11581 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
    11641 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
    11701 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
    11761 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
    11821 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
    11881 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
    11941 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
    12001 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
    12061 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
    12121 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
    12181 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
    12241 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
    12301 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
    12361 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
    12421 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
    12481 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
    12541 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
    12601 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
    12661 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
    12721 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
    12781 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
    12841 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
    12901 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
    12961 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
    13021 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
    13081 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
    13141 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
    13201 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
    13261 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
    13321 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
    13381 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
    13441 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
    13501 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
    13561 ccgcgcacat ttccccgaaa agtgccacct gac
//
