LOCUS       sequence-237476-       13027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  U6-sgRNA EFS-ProCas9-Flavi-P2A-Puro.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13027)
  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 13027)
  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: pCF704
FEATURES             Location/Qualifiers
     source          1..13027
                     /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        2608..2856
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2608..2628
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2779..2798
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        3031..3242
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3250..3259
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3262..3282
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4300..6798
                     /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             7420..7440
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7450..7506
                     /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             7507..8103
                     /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     8000..8020
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8119..8707
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8172..8192)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8590..8601)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8779..9012
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9038..9055)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9044..9268
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9314..9742
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9401..9420)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9611..9632
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9751..9771)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9756..10085
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9936..10071
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9998..10017
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10133..10180
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10199..10573
                     /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    10703..10824
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10740..10759)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10794..10813
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10873..10889)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10873..10889)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10886..10908)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10897..10913
                     /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(10921..10927,10928..10945,10946..10951))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10966..10987
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11104..11121)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11275..11863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11355..11374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12034..12825,12826..12894))
                     /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     12657..12676
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12895..12999)
                     /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 aggtaccgag ggcctatttc ccatgattcc ttcatatttg
     2641 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
     2701 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     2761 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     2821 ttggctttat atatcttgtg gaaaggacga aacaccggag acggaggacg acgaacgtct
     2881 ctgtttaaga gctatgctgg aaacagcata gcaagtttaa ataaggctag tccgttatca
     2941 acttgaaaaa gtggcaccga gtcggtgctt tttttgaatt cgctagctag gtcttgaaag
     3001 gagtgggaat tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
     3061 agaagttggg gggaggggtc ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa
     3121 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
     3181 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
     3241 aggaccggtg ccaccatggc cccaaagaag aagcggaagg tcagcgctaa ccccatcaac
     3301 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
     3361 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt
     3421 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
     3481 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
     3541 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     3601 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     3661 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     3721 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     3781 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     3841 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     3901 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt
     3961 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     4021 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     4081 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     4141 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     4201 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     4261 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     4321 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     4381 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     4441 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     4501 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     4561 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     4621 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
     4681 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     4741 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     4801 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag
     4861 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag
     4921 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa
     4981 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag
     5041 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa
     5101 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg
     5161 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg
     5221 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag
     5281 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac
     5341 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag
     5401 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg
     5461 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag
     5521 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg
     5581 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac
     5641 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct
     5701 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc
     5761 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc
     5821 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg
     5881 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc
     5941 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca
     6001 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga
     6061 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct
     6121 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag
     6181 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg
     6241 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc
     6301 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag
     6361 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat
     6421 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac
     6481 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg
     6541 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc
     6601 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc
     6661 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg
     6721 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg
     6781 tctcagctgg gaggcgacaa acaaaagaaa cgcggaggca aggacaagaa gtacagcatc
     6841 ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg
     6901 cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg
     6961 atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc
     7021 gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc
     7081 aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg
     7141 gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc
     7201 taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac
     7261 aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac
     7321 ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag
     7381 ctggtgcaga cctacaacca gctgttcgag gaaactagtc caaaaaagaa gagaaaggta
     7441 ggatccggcg caacaaactt ctctctgctg aaacaagccg gagatgtcga agagaatcct
     7501 ggaccgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     7561 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     7621 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     7681 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     7741 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     7801 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     7861 gagcccgcgt ggttcctggc caccgtcgga gtctcgcccg accaccaggg caagggtctg
     7921 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     7981 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     8041 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     8101 tgaacgcgtt aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     8161 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     8221 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     8281 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     8341 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     8401 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     8461 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     8521 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     8581 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     8641 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     8701 tccccgcgtc gactttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     8761 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
     8821 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     8881 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     8941 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     9001 aaatctctag cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     9061 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     9121 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     9181 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     9241 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     9301 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     9361 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     9421 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     9481 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     9541 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     9601 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     9661 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     9721 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     9781 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     9841 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     9901 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     9961 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
    10021 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
    10081 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa
    10141 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
    10201 ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga
    10261 gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt
    10321 ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa
    10381 caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt
    10441 cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc
    10501 gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga
    10561 ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt
    10621 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
    10681 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
    10741 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
    10801 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
    10861 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
    10921 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
    10981 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
    11041 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
    11101 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
    11161 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
    11221 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
    11281 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
    11341 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
    11401 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
    11461 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
    11521 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
    11581 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
    11641 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
    11701 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
    11761 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
    11821 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
    11881 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
    11941 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
    12001 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
    12061 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
    12121 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
    12181 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
    12241 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
    12301 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
    12361 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
    12421 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
    12481 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
    12541 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
    12601 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
    12661 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
    12721 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
    12781 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
    12841 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
    12901 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
    12961 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
    13021 acctgac
//
