LOCUS       sequence-275142-       13400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentiviral gRNA plasmid targeting human CDKN1A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13400)
  AUTHORS   Kwan SY, Sheel A, Song CQ, Zhang XO, Jiang T, Dang H, Cao Y, Ozata 
            DM, Mou H, Yin H, Weng Z, Wang XW, Xue W
  TITLE     Depletion of TRRAP induces p53-independent senescence in liver 
            cancer by downregulating mitotic genes.
  JOURNAL   Hepatology. 2019 Jun 12. doi: 10.1002/hep.30807.
  PUBMED    31188495
REFERENCE   2  (bases 1 to 13400)
  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: "Hepatology.
            2019 Jun 12. doi: 10.1002/hep.30807."
COMMENT     Sequence Label: CDKN1A sgRNA
FEATURES             Location/Qualifiers
     source          1..13400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..126
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    619..852
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1037..1081
                     /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    1379..1496
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1547..1795
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1547..1567
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1718..1737
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1816..1891
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1953..2164
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2183..2192
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2189..6292
                     /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             2189..6292
                     /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             6293..6340
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6341..6364
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6374..6430
                     /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"
     misc_feature    7427..8015
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7480..7500)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7898..7909)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8087..8320
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8346..8363)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8352..8576
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8622..9050
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8709..8728)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8919..8940
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9059..9079)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9064..9393
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9244..9379
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9306..9325
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9441..9488
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9507..9881
                     /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    10011..10132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10048..10067)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10102..10121
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10181..10197)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10181..10197)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10194..10216)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10205..10221
                     /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(10229..10235,10236..10253,10254..10259))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10274..10295
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10412..10429)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10583..11171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10663..10682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11342..12133,12134..12202))
                     /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     11965..11984
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12203..12307)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12382..12401)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        12577..12956
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        12958..13156
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     13111..13131
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             13174..13354
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga
       61 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag
      121 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca
      181 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga
      241 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
      301 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
      361 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
      421 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag
      481 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
      541 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
      601 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
      661 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt
      721 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
      781 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga
      841 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
      901 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat
      961 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
     1021 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
     1081 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
     1141 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
     1201 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag
     1261 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt
     1321 gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt
     1381 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
     1441 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg
     1501 tttattacag ggacagcaga gatccagttt ggttaattaa ggtaccgagg gcctatttcc
     1561 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa
     1621 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
     1681 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac
     1741 ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgtcac
     1801 cgagacacca ctggagtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     1861 tcaacttgaa aaagtggcac cgagtcggtg cttttttgaa ttcgctagct aggtcttgaa
     1921 aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
     1981 cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt ggcgcggggt
     2041 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
     2101 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
     2161 acaggaccgg ttctagagcg ctgccaccat ggacaagaag tacagcatcg gcctggacat
     2221 cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa
     2281 attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct
     2341 gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag
     2401 atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc
     2461 caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa
     2521 gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa
     2581 gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct
     2641 gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga
     2701 gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac
     2761 ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat
     2821 cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg
     2881 cgagaagaag aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa
     2941 cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta
     3001 cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct
     3061 ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga
     3121 gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga
     3181 cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt
     3241 cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga
     3301 gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt
     3361 gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc
     3421 ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc
     3481 attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta
     3541 cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga
     3601 aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt
     3661 catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca
     3721 cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac
     3781 cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct
     3841 gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa
     3901 aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct
     3961 gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga
     4021 aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat
     4081 gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct
     4141 gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg
     4201 ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag
     4261 aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc
     4321 ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc
     4381 cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat
     4441 gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca
     4501 gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct
     4561 gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct
     4621 gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa
     4681 ccggctgtcc gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc
     4741 catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc
     4801 ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct
     4861 gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact
     4921 ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt
     4981 ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg
     5041 ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca
     5101 gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc
     5161 cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg
     5221 cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa
     5281 ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac
     5341 cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga
     5401 gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca
     5461 agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct
     5521 gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta
     5581 cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa
     5641 gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag
     5701 aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa
     5761 aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa
     5821 gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa
     5881 atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga
     5941 taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga
     6001 gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct
     6061 gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca
     6121 cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat
     6181 cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag
     6241 catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acaagcgacc
     6301 tgccgccaca aagaaggctg gacaggctaa gaagaagaaa gattacaaag acgatgacga
     6361 taagggatcc ggcgcaacaa acttctctct gctgaaacaa gccggagatg tcgaagagaa
     6421 tcctggaccg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     6481 gttcgacagc gtgtccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     6541 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     6601 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
     6661 tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     6721 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     6781 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     6841 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     6901 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     6961 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     7021 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     7081 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     7141 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     7201 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     7261 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     7321 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac
     7381 cgatggctgt gtagaagtac ttcgccgata gacgcgttaa gtcgacaatc aacctctgga
     7441 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     7501 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     7561 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     7621 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     7681 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     7741 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     7801 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     7861 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     7921 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     7981 gacgagtcgg atctcccttt gggccgcctc cccgcgtcga ctttaagacc aatgacttac
     8041 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     8101 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
     8161 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     8221 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     8281 tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt aaacccgctg
     8341 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     8401 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     8461 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     8521 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     8581 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     8641 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     8701 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     8761 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     8821 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     8881 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     8941 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     9001 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     9061 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     9121 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     9181 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     9241 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     9301 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     9361 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     9421 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
     9481 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca
     9541 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
     9601 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
     9661 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
     9721 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
     9781 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
     9841 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg
     9901 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     9961 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
    10021 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
    10081 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
    10141 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
    10201 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
    10261 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
    10321 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
    10381 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
    10441 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
    10501 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
    10561 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
    10621 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
    10681 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
    10741 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
    10801 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
    10861 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
    10921 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
    10981 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
    11041 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
    11101 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
    11161 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
    11221 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
    11281 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
    11341 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
    11401 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
    11461 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
    11521 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
    11581 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
    11641 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
    11701 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
    11761 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
    11821 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
    11881 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
    11941 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
    12001 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
    12061 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
    12121 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
    12181 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
    12241 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
    12301 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
    12361 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
    12421 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
    12481 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
    12541 gcttcgcgat gtacgggcca gatatacgcg cgcgttgaca ttgattattg actagttatt
    12601 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
    12661 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
    12721 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
    12781 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
    12841 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
    12901 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
    12961 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
    13021 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
    13081 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
    13141 aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga
    13201 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
    13261 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
    13321 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
    13381 cgaaagggaa accagaggag
//
