LOCUS       sequence-438023-       13212 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of sgRNA targeting the locus of human PTPN14.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13212)
  AUTHORS   Blakely WJ, Hatterschide J, White EA
  TITLE     HPV18 E7 inhibits LATS1 kinase and activates YAP1 by degrading 
            PTPN14.
  JOURNAL   mBio. 2024 Sep 9:e0181124. doi: 10.1128/mbio.01811-24.
  PUBMED    39248565
REFERENCE   2  (bases 1 to 13212)
  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: "mBio. 2024 
            Sep 9:e0181124. doi: 10.1128/mbio.01811-24."
COMMENT     Sequence Label: p8391 LentiCRISPRv2 Neo sgPTPN14-1
FEATURES             Location/Qualifiers
     source          1..13212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..171
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             214..394
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    441..566
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1059..1292
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1477..1521
                     /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    1819..1936
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1987..2235
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1987..2007
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2158..2177
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2257..2332
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2394..2605
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2624..2633
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2630..6733
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2630..6733
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6734..6781
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6782..6805
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6815..6871
                     /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             6872..7663
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6923..6942)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7533..7552
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    7679..8267
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7732..7752)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8150..8161)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8339..8572
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8598..8615)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8604..8828
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8874..9302
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8961..8980)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9171..9192
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9311..9331)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9316..9645
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9496..9631
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9558..9577
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9693..9740
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9759..10133
                     /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    10263..10384
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10300..10319)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10354..10373
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10433..10449)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10433..10449)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10446..10468)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10457..10473
                     /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(10481..10487,10488..10505,10506..10511))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10526..10547
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10664..10681)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10835..11423)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10915..10934)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11594..12385,12386..12454))
                     /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     12217..12236
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12455..12559)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12634..12653)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        12829..13208
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(13210..13212,1..196)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
       61 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      121 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      181 aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga
      241 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      301 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      361 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      421 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      481 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      541 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      601 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      661 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      721 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
      781 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
      841 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
      901 aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
      961 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     1021 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     1081 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
     1141 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     1201 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     1261 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     1321 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     1381 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
     1441 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
     1501 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
     1561 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
     1621 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
     1681 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
     1741 gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat
     1801 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
     1861 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
     1921 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggttaattaa
     1981 ggtaccgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     2041 agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     2101 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     2161 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     2221 aaaggacgaa acaccgcatg actgtctcat aatcgggttt tagagctaga aatagcaagt
     2281 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttga
     2341 attcgctagc taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga
     2401 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gatccggtgc
     2461 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     2521 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     2581 caacgggttt gccgccagaa cacaggaccg gttctagagc gctgccacca tggacaagaa
     2641 gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga
     2701 gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa
     2761 gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct
     2821 gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga
     2881 gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc
     2941 cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga
     3001 cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga
     3061 cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt
     3121 ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct
     3181 gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag
     3241 cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa
     3301 tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct
     3361 gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact
     3421 gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga
     3481 ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga
     3541 catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag
     3601 atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc
     3661 tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga
     3721 cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga
     3781 cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac
     3841 cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg
     3901 gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct
     3961 gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg
     4021 gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa
     4081 gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa
     4141 cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct
     4201 gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca
     4261 gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct
     4321 gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga
     4381 agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa
     4441 ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac
     4501 actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga
     4561 cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg
     4621 gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa
     4681 gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt
     4741 taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat
     4801 tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt
     4861 ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc
     4921 cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat
     4981 cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac
     5041 ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt
     5101 ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca
     5161 gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg
     5221 gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg
     5281 gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga
     5341 gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac
     5401 ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga
     5461 cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc
     5521 cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc
     5581 ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct
     5641 ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa
     5701 gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa
     5761 ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga
     5821 gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg
     5881 gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg
     5941 cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa
     6001 ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct
     6061 ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct
     6121 ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc
     6181 caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt
     6241 cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa
     6301 cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa
     6361 gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca
     6421 ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga
     6481 cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga
     6541 gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt
     6601 caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga
     6661 cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca
     6721 gctgggaggc gacaagcgac ctgccgccac aaagaaggct ggacaggcta agaagaagaa
     6781 agattacaaa gacgatgacg ataagggatc cggcgcaaca aacttctctc tgctgaaaca
     6841 agccggagat gtcgaagaga atcctggacc gattgaacaa gatggattgc acgcaggttc
     6901 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     6961 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     7021 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     7081 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     7141 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     7201 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     7261 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     7321 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     7381 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     7441 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     7501 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     7561 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     7621 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgaacgcgtt aagtcgacaa
     7681 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     7741 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     7801 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     7861 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     7921 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     7981 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     8041 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     8101 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     8161 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     8221 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc gactttaaga
     8281 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     8341 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
     8401 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     8461 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     8521 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagggcccgt
     8581 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     8641 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     8701 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     8761 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     8821 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     8881 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     8941 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     9001 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     9061 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     9121 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     9181 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     9241 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     9301 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     9361 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     9421 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     9481 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     9541 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     9601 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     9661 tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata
     9721 tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg
     9781 ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg
     9841 ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc
     9901 tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg
     9961 tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg
    10021 acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc
    10081 tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc
    10141 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
    10201 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
    10261 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
    10321 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
    10381 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
    10441 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
    10501 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
    10561 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
    10621 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
    10681 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
    10741 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
    10801 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
    10861 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
    10921 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
    10981 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
    11041 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
    11101 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
    11161 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
    11221 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
    11281 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
    11341 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
    11401 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
    11461 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
    11521 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
    11581 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
    11641 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
    11701 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
    11761 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
    11821 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
    11881 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
    11941 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
    12001 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
    12061 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
    12121 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
    12181 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
    12241 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
    12301 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
    12361 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
    12421 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
    12481 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
    12541 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
    12601 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
    12661 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
    12721 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
    12781 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgcgcgttga cattgattat
    12841 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
    12901 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
    12961 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
    13021 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
    13081 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
    13141 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
    13201 ttaccatggt ga
//
