LOCUS       sequence-261775-       14608 bp    DNA     circular SYN 15-APR-2025
DEFINITION  EGFP reporter sequence was removed from TLCV2 vector and two gRNAs 
            targeting mouse Exoc7 gene were cloned into the modified TLCV2 
            vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14608)
  AUTHORS   Wang S, Crisman L, Miller J, Datta I, Gulbranson DR, Tian Y, Yin Q, 
            Yu H, Shen J
  TITLE     Inducible Exoc7/Exo70 knockout reveals a critical role of the 
            exocyst in insulin-regulated GLUT4 exocytosis.
  JOURNAL   J Biol Chem. 2019 Dec 27;294(52):19988-19996. doi: 
            10.1074/jbc.RA119.010821. Epub 2019 Nov 18.
  PUBMED    31740584
REFERENCE   2  (bases 1 to 14608)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1074/jbc.RA119.010821"; journalName: "J Biol Chem"; date: 
            "2019-12-27- 27"; volume: "294"; issue: "52"; pages: "19988-19996"
COMMENT     Sequence Label: TLCV2-Exoc7-Ex5&10
FEATURES             Location/Qualifiers
     source          1..14608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    36..54
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    71..89
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    107..125
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    143..161
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    178..196
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    214..232
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     279..303
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      332..341
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             338..4441
                     /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             338..4441
                     /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             4442..4489
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4490..4513
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4523..4576
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     promoter        4590..4801
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             4823..5419
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4823..4842)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5319..5339
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             5429..5485
                     /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             join(5486..6103,6104..6109,6110..6145,6146..6148,6149..6184,
                     6185..6187,6188..6223,6224..6232)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    6253..6841
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6306..6326)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6724..6735)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6913..7146
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7172..7189)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7178..7402
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7448..7876
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7535..7554)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7745..7766
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7885..7905)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7890..8219
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8070..8205
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8132..8151
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8267..8314
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8333..8707
                     /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    8837..8958
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8874..8893)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8928..8947
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9007..9023)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9007..9023)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9020..9042)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9031..9047
                     /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(9055..9061,9062..9079,9080..9085))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9100..9121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9238..9255)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9409..9997)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9489..9508)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10168..10959,10960..11028))
                     /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     10791..10810
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11029..11133)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(11208..11227)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        11399..11778
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11780..11978
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             11996..12176
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    12223..12348
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12841..13074
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             13259..13303
                     /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    13601..13718
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        complement(13791..13866)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(13889..14202)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     complement(13950..13970)
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     promoter        14218..14466
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     14218..14238
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     14389..14408
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        14488..14563
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        join(14600..14608,1..306)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    join(14608,1..18)
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgatg
       61 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
      121 agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttatcc
      181 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
      241 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
      301 atcgcctgga gaataccggt tctagagcgc tgccaccatg gacaagaagt acagcatcgg
      361 cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc
      421 cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat
      481 cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc
      541 cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa
      601 cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga
      661 agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta
      721 ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa
      781 ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt
      841 cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct
      901 ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc
      961 caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca
     1021 gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct
     1081 gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa
     1141 ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga
     1201 cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt
     1261 gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca
     1321 ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa
     1381 agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag
     1441 ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga
     1501 actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg
     1561 cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga
     1621 tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat
     1681 cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa
     1741 gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc
     1801 ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct
     1861 gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa
     1921 atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat
     1981 cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta
     2041 cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa
     2101 cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga
     2161 caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga
     2221 cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat
     2281 gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa
     2341 cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt
     2401 cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat
     2461 ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc
     2521 cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt
     2581 gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca
     2641 gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat
     2701 caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa
     2761 cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact
     2821 ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa
     2881 ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga
     2941 caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa
     3001 cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct
     3061 gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac
     3121 aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa
     3181 gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa
     3241 ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta
     3301 cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt
     3361 cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga
     3421 aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac
     3481 cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga
     3541 aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag
     3601 catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga
     3661 gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc
     3721 taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa
     3781 agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat
     3841 catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa
     3901 agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa
     3961 cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct
     4021 gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc
     4081 ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga
     4141 gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga
     4201 caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa
     4261 tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga
     4321 caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat
     4381 ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga
     4441 caagcgacct gccgccacaa agaaggctgg acaggctaag aagaagaaag attacaaaga
     4501 cgatgacgat aagggatccg gagagggcag aggaagtctg ctaacatgcg gtgacgtcga
     4561 ggagaatcct ggcccagcta gctaaatccg ggcagagcgc acatcgccca cagtccccga
     4621 gaagttgggg ggaggggtcg gcaattgatc cggtgcctag agaaggtggc gcggggtaaa
     4681 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     4741 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     4801 gactgcgatc gcaatgtaca gtatgaccga gtacaagccc acggtgcgcc tcgccacccg
     4861 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     4921 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     4981 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     5041 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     5101 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     5161 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggag tctcgcccga
     5221 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     5281 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     5341 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     5401 gacccgcaag cccggtgccg gttccggcgc aacaaacttc tctctgctga aacaagccgg
     5461 agatgtcgaa gagaatcctg gaccgatgtc tagactggac aagagcaaag tcataaacgg
     5521 agctctggaa ttactcaatg gtgtcggtat cgaaggcctg acgacaagga aactcgctca
     5581 aaagctggga gttgagcagc ctaccctgta ctggcacgtg aagaacaagc gggccctgct
     5641 cgatgccctg ccaatcgaga tgctggacag gcatcatacc cacttctgcc ccctggaagg
     5701 cgagtcatgg caagactttc tgcggaacaa cgccaagtca taccgctgtg ctctcctctc
     5761 acatcgcgac ggggctaaag tgcatctcgg cacccgccca acagagaaac agtacgaaac
     5821 cctggaaaat cagctcgcgt tcctgtgtca gcaaggcttc tccctggaga acgcactgta
     5881 cgctctgtcc gccgtgggcc actttacact gggctgcgta ttggaggaac aggagcatca
     5941 agtagcaaaa gaggaaagag agacacctac caccgattct atgcccccac ttctgagaca
     6001 agcaattgag ctgttcgacc ggcagggagc cgaacctgcc ttccttttcg gcctggaact
     6061 aatcatatgt ggcctggaga aacagctaaa gtgcgaaagc ggcgggccga ccgacgccct
     6121 tgacgatttt gacttagaca tgctcccagc cgatgccctt gacgactttg accttgatat
     6181 gctgcctgct gacgctcttg acgattttga ccttgacatg ctccccgggt aactcgagcg
     6241 cgttaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     6301 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     6361 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     6421 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     6481 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     6541 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     6601 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     6661 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     6721 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     6781 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     6841 cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     6901 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt
     6961 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     7021 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     7081 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     7141 ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     7201 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     7261 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     7321 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     7381 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     7441 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     7501 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     7561 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     7621 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     7681 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     7741 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     7801 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     7861 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     7921 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     7981 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     8041 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     8101 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     8161 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     8221 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc
     8281 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa
     8341 gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg
     8401 gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg
     8461 ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct
     8521 ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc
     8581 cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg
     8641 gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca
     8701 ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     8761 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     8821 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     8881 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     8941 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     9001 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     9061 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     9121 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     9181 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     9241 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     9301 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     9361 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     9421 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     9481 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     9541 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     9601 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     9661 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     9721 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     9781 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     9841 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     9901 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     9961 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
    10021 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
    10081 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
    10141 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
    10201 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
    10261 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
    10321 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
    10381 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
    10441 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
    10501 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
    10561 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
    10621 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
    10681 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
    10741 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
    10801 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
    10861 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
    10921 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
    10981 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
    11041 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
    11101 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
    11161 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
    11221 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
    11281 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
    11341 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
    11401 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
    11461 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
    11521 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
    11581 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
    11641 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
    11701 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
    11761 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
    11821 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
    11881 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
    11941 ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc
    12001 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
    12061 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
    12121 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
    12181 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
    12241 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
    12301 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
    12361 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
    12421 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
    12481 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
    12541 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
    12601 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
    12661 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
    12721 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
    12781 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
    12841 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
    12901 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
    12961 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
    13021 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
    13081 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
    13141 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
    13201 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
    13261 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
    13321 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
    13381 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
    13441 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
    13501 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg
    13561 tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
    13621 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
    13681 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
    13741 gagatccagt ttggttaatt aaggtaccgc ggcgtctcag tacaaaaaaa gcaccgactc
    13801 ggtgccactt tttcaagttg ataacggact agccttattt taacttgcta tttctagctc
    13861 taaaactcgc tcaaacagca gcttctcaaa caaggctttt ctccaaggga tatttatagt
    13921 ctcaaaacac acaattactt tacagttagg gtgagtttcc ttttgtgctg ttttttaaaa
    13981 taataattta gtatttgtat ctcttataga aatccaagcc tatcatgtaa aatgtagcta
    14041 gtattaaaaa gaacagatta tctgtctttt atcgcacatt aagcctctat agttactagg
    14101 aaatattata tgcaaattaa ccggggcagg ggagtagccg agcttctccc acaagtctgt
    14161 gcgagggggc cggcgcgggc ctagagatgg cggcgtcgga tctataccgg tggtaccgag
    14221 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
    14281 attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
    14341 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
    14401 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
    14461 aacaccgttc tagagctttg gccccaagtt ttagagctag aaatagcaag ttaaaataag
    14521 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg aattcacttt
    14581 ggccgcgaat cgatatgtcg agtttact
//
