LOCUS       sequence-298911-       14234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hu6 HGPS sgRNA expression and ABE7.10max VRQR lentiviral plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14234)
  AUTHORS   Koblan LW, Erdos MR, Wilson C, Cabral WA, Levy JM, Xiong ZM, Tavarez
            UL, Davison LM, Gete YG, Mao X, Newby GA, Doherty SP, Narisu N, 
            Sheng Q, Krilow C, Lin CY, Gordon LB, Cao K, Collins FS, Brown JD, 
            Liu DR
  TITLE     In vivo base editing rescues Hutchinson-Gilford progeria syndrome in
            mice.
  JOURNAL   Nature. 2021 Jan;589(7843):608-614. doi: 10.1038/s41586-020-03086-7.
            Epub 2021 Jan 6.
  PUBMED    33408413
REFERENCE   2  (bases 1 to 14234)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41586-020-03086-7"; journalName: "Nature"; date: 
            "2021-01"; volume: "589"; issue: "7843"; pages: "608-614"
COMMENT     Sequence Label: hu6 HGPS sgRNA expression and ABE7.10max VRQR
            ...
FEATURES             Location/Qualifiers
     source          1..14234
                     /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"
     misc_RNA        2877..2952
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3014..3225
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3275..3295
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4484..8584
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8627..8647
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8657..8713
                     /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             8714..9310
                     /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     9207..9227
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9326..9914
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9379..9399)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9797..9808)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9986..10219
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10245..10262)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10251..10475
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10521..10949
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10608..10627)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10818..10839
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10958..10978)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10963..11292
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11143..11278
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11205..11224
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11340..11387
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11406..11780
                     /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    11910..12031
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11947..11966)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12001..12020
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12080..12096)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12080..12096)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12093..12115)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12104..12120
                     /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(12128..12134,12135..12152,12153..12158))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12173..12194
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12311..12328)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12482..13070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12562..12581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13241..14032,14033..14101))
                     /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     13864..13883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14102..14206)
                     /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 aacaccggtc cacccacctg ggctccgttt
     2881 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
     2941 ccgagtcggt gcttttttga attcgctagc taggtcttga aaggagtggg aattggctcc
     3001 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     3061 gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     3121 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     3181 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggaccg gtgccaccat
     3241 gaaacggaca gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtctctga
     3301 agtcgagttt agccacgagt attggatgag gcacgcactg accctggcaa agcgagcatg
     3361 ggatgaaaga gaagtccccg tgggcgccgt gctggtgcac aacaatagag tgatcggaga
     3421 gggatggaac aggccaatcg gccgccacga ccctaccgca cacgcagaga tcatggcact
     3481 gaggcaggga ggcctggtca tgcagaatta ccgcctgatc gatgccaccc tgtatgtgac
     3541 actggagcca tgcgtgatgt gcgcaggagc aatgatccac agcaggatcg gaagagtggt
     3601 gttcggagca cgggacgcca agaccggcgc agcaggctcc ctgatggatg tgctgcacca
     3661 ccccggcatg aaccaccggg tggagatcac agagggaatc ctggcagacg agtgcgccgc
     3721 cctgctgagc gatttcttta gaatgcggag acaggagatc aaggcccaga agaaggcaca
     3781 gagctccacc gactctggag gatctagcgg aggatcctct ggaagcgaga caccaggcac
     3841 aagcgagtcc gccacaccag agagctccgg cggctcctcc ggaggatcct ctgaggtgga
     3901 gttttcccac gagtactgga tgagacatgc cctgaccctg gccaagaggg cacgcgatga
     3961 gagggaggtg cctgtgggag ccgtgctggt gctgaacaat agagtgatcg gcgagggctg
     4021 gaacagagcc atcggcctgc acgacccaac agcccatgcc gaaattatgg ccctgagaca
     4081 gggcggcctg gtcatgcaga actacagact gattgacgcc accctgtacg tgacattcga
     4141 gccttgcgtg atgtgcgccg gcgccatgat ccactctagg atcggccgcg tggtgtttgg
     4201 cgtgaggaac gcaaaaaccg gcgccgcagg ctccctgatg gacgtgctgc actaccccgg
     4261 catgaatcac cgcgtcgaaa ttaccgaggg aatcctggca gatgaatgtg ccgccctgct
     4321 gtgctatttc tttcggatgc ctagacaggt gttcaatgct cagaagaagg cccagagctc
     4381 caccgactcc ggaggatcta gcggaggctc ctctggctct gagacacctg gcacaagcga
     4441 gagcgcaaca cctgaaagca gcgggggcag cagcgggggg tcagacaaga agtacagcat
     4501 cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt
     4561 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct
     4621 gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac
     4681 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag
     4741 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt
     4801 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc
     4861 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga
     4921 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca
     4981 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca
     5041 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga
     5101 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc
     5161 ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg
     5221 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag
     5281 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc
     5341 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag
     5401 agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga
     5461 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta
     5521 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc
     5581 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga
     5641 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa
     5701 cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga
     5761 agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg
     5821 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag
     5881 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc
     5941 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt
     6001 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt
     6061 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc
     6121 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga
     6181 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt
     6241 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct
     6301 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga
     6361 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt
     6421 gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat
     6481 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg
     6541 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga
     6601 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct
     6661 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct
     6721 cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa
     6781 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg
     6841 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca
     6901 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga
     6961 actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct
     7021 gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag
     7081 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct
     7141 gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg
     7201 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat
     7261 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga
     7321 caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg
     7381 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc
     7441 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga
     7501 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca
     7561 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa
     7621 gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg
     7681 cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct
     7741 gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa
     7801 agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga
     7861 ccctaagaag tacggcggct tcgtgagccc caccgtggcc tattctgtgc tggtggtggc
     7921 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac
     7981 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta
     8041 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga
     8101 aaacggccgg aagagaatgc tggcctcagc cagagaactg cagaagggaa acgaactggc
     8161 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg
     8221 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga
     8281 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct
     8341 ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga
     8401 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt
     8461 tgacaccacc atcgaccgga agcagtacag aagcaccaaa gaggtgctgg acgccaccct
     8521 gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg
     8581 tgactctggc ggctcaaaaa gaaccgccga cggcagcgaa ttcgagccca agaagaagag
     8641 gaaagtcgga tccggggcca ccaacttttc tctgctgaaa caagccggag atgtcgaaga
     8701 gaatcctgga ccgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     8761 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     8821 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     8881 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     8941 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     9001 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     9061 gcccaaggag cccgcgtggt tcctggccac cgtcggagtc tcgcccgacc accagggcaa
     9121 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     9181 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     9241 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     9301 cggtgcctga acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     9361 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     9421 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     9481 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     9541 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     9601 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     9661 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     9721 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     9781 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     9841 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     9901 ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc
     9961 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
    10021 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
    10081 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
    10141 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
    10201 gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
    10261 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
    10321 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
    10381 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
    10441 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
    10501 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
    10561 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
    10621 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
    10681 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
    10741 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
    10801 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
    10861 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
    10921 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
    10981 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
    11041 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
    11101 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
    11161 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
    11221 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
    11281 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
    11341 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
    11401 aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag
    11461 cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg
    11521 ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc
    11581 cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt
    11641 cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg
    11701 agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg
    11761 tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg
    11821 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
    11881 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
    11941 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
    12001 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
    12061 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
    12121 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
    12181 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
    12241 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
    12301 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
    12361 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
    12421 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
    12481 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
    12541 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
    12601 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
    12661 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
    12721 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
    12781 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
    12841 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
    12901 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
    12961 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
    13021 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
    13081 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
    13141 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
    13201 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
    13261 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
    13321 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
    13381 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
    13441 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
    13501 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
    13561 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
    13621 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
    13681 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
    13741 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
    13801 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
    13861 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
    13921 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
    13981 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
    14041 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
    14101 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
    14161 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
    14221 aagtgccacc tgac
//
