LOCUS       40924_3286       13069 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Dox-inducible CRISPR interference (CRISPRi) knock in construct into 
            the AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13069)
  AUTHORS   Mandegar MA, Huebsch N, Frolov EB, Shin E, Truong A, Olvera MP, Chan
            AH, Miyaoka Y, Holmes K, Spencer CI, Judge LM, Gordon DE, Eskildsen 
            TV, Villalta JE, Horlbeck MA, Gilbert LA, Krogan NJ, Sheikh SP, 
            Weissman JS, Qi LS, So PL, Conklin BR
  TITLE     CRISPR Interference Efficiently Induces Specific and Reversible Gene
            Silencing in Human iPSCs.
  JOURNAL   Cell Stem Cell. 2016 Apr 7;18(4):541-53. doi: 
            10.1016/j.stem.2016.01.022. Epub 2016 Mar 10.
  PUBMED    26971820
REFERENCE   2  (bases 1 to 13069)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 13069)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.stem.2016.01.022"; journalName: "Cell Stem Cell"; date: 
            "2016-04-7- 7"; volume: "18"; issue: "4"; pages: "541-53"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..13069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             54..911
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1085..1673
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(1721..2557)
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        2583..2960
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     regulatory      2977..2986
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3013..3198
                     /codon_start=1
                     /label=KRAB
                     /note="Kruppel-associated box (KRAB) transcriptional
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             3220..7323
                     /codon_start=1
                     /label=dCas9
                     /note="catalytically dead mutant of the Cas9 endonuclease
                     from the Streptococcus pyogenes Type II CRISPR/Cas system"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7327..7353
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7372..7392
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7399..7419
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7493..7626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7663..7679)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7663..7679)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7676..7698)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7687..7703
                     /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(7711..7741)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7756..7777)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(7894..7911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7967..8000
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     8331..8350
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also
                     called rb-glob-pA-term-R"
     polyA_signal    complement(8349..8404)
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     8450..8469
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(8562..9305)
                     /codon_start=1
                     /label=Tet-On(R) 3G
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(9337..9356)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids,
                     forward primer"
     intron          complement(9364..10381)
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     promoter        complement(10383..10660)
                     /label=chicken beta-actin promoter
     enhancer        10662..11041
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    complement(11047..11271)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(11300..12100)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(12122..12175)
                     /codon_start=1
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    complement(12199..12224)
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    12231..12264
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    complement(12266..13069)
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
ORIGIN
        1 aagggcgaat tcaaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
       61 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      121 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      181 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      241 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      301 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
      361 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
      421 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
      481 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
      541 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
      601 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
      661 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
      721 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
      781 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
      841 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
      901 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
      961 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1021 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1081 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1141 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1201 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     1261 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1321 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1381 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1441 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1501 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1561 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1621 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     1681 gcaacgcggc ctttttacgg ttcctggcga attcgccctt agagcagagc caggaacccc
     1741 tgtagggaag gggcaggaga gccaggggca tgagatggtg gacgaggaag ggggacaggg
     1801 aagcctgagc gcctctcctg ggcttgccaa ggactcaaac ccagaagccc agagcagggc
     1861 cttagggaag cgggaccctg ctctgggcgg aggaatatgt cccagatagc actggggact
     1921 ctttaaggaa agaaggatgg agaaagagaa agggagtaga ggcggccacg acctggtgaa
     1981 cacctaggac gcaccattct cacaaaggga gttttccaca cggacacccc cctcctcacc
     2041 acagccctgc caggacgggg ctggctactg gccttatctc acaggtaaaa ctgacgcacg
     2101 gaggaacaat ataaattggg gactagaaag gtgaagagcc aaagttagaa ctcaggacca
     2161 acttattctg attttgtttt tccaaactgc ttctcctctt gggaagtgta aggaagctgc
     2221 agcaccagga tcagtgaaac gcaccagaca gccgcgtcag agcagctcag gttctgggag
     2281 agggtagcgc agggtggcca ctgagaaccg ggcaggtcac gcatcccccc cttccctccc
     2341 accccctgcc aagctctccc tcccaggatc ctctctggct ccatcgtaag caaaccttag
     2401 aggttctggc aaggagagag atggctccag gaaatggggg tgtgtcacca gataaggaat
     2461 ctgcctaaca ggaggtgggg gttagaccca atatcaggag actaggaagg aggaggccta
     2521 aggatggggc ttttctgtca ccaatcctgt ccctagtaaa gctgatatct ccaccccaca
     2581 gtggtttact ccctatcagt gatagagaac gtatgaagag tttactccct atcagtgata
     2641 gagaacgtat gcagacttta ctccctatca gtgatagaga acgtataagg agtttactcc
     2701 ctatcagtga tagagaacgt atgaccagtt tactccctat cagtgataga gaacgtatct
     2761 acagtttact ccctatcagt gatagagaac gtatatccag tttactccct atcagtgata
     2821 gagaacgtat aagctttagg cgtgtacggt gggcgcctat aaaagcagag ctcgtttagt
     2881 gaaccgtcag atcgcctgga gcaattccac aacacttttg tcttatacca actttccgta
     2941 ccacttccta ccctcgtaaa cttaaggtta attaacgcca ccatggatgc taagtcacta
     3001 actgcctggt cccggacact ggtgaccttc aaggatgtat ttgtggactt caccagggag
     3061 gagtggaagc tgctggacac tgctcagcag atcgtgtaca gaaatgtgat gctggagaac
     3121 tataagaacc tggtttcctt gggttatcag cttactaagc cagatgtgat cctccggttg
     3181 gagaagggag aagagcccgg cggttccggc ggagggtcga tggacaagaa gtacagcatc
     3241 ggcctggcca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg
     3301 cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg
     3361 atcggcgccc tgctgttcga cagcggagaa acagccgagg ccacccggct gaagagaacc
     3421 gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc
     3481 aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg
     3541 gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc
     3601 taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac
     3661 aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac
     3721 ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag
     3781 ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac
     3841 gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc
     3901 cagctgcccg gcgagaagaa gaatggcctg ttcggcaacc tgattgccct gagcctgggc
     3961 ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc
     4021 aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc
     4081 gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga
     4141 gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag
     4201 caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac
     4261 aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacatcga tggcggagcc
     4321 agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag
     4381 gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac
     4441 ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa
     4501 gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc
     4561 atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga
     4621 aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgccagc
     4681 gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg
     4741 ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt acaacgagct gaccaaagtg
     4801 aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaagcc
     4861 atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac
     4921 tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc
     4981 aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg
     5041 gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag
     5101 gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg
     5161 atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc
     5221 aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc
     5281 ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac
     5341 atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg
     5401 gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc
     5461 gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac
     5521 cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc
     5581 atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag
     5641 aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa
     5701 ctggacatca accggctgtc cgactacgat gtggacgcta tcgtgcctca gagctttctg
     5761 aaggacgact ccatcgataa caaagtgctg actcggagcg acaagaaccg gggcaagagc
     5821 gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg ccagctgctg
     5881 aatgccaagc tgattaccca gaggaagttc gacaatctga ccaaggccga gagaggcggc
     5941 ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc
     6001 acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaacgac
     6061 aaactgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg
     6121 aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc
     6181 tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag
     6241 ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag
     6301 gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag
     6361 accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc
     6421 gaaacaggcg agatcgtgtg ggataagggc cgggactttg ccaccgtgcg gaaagtgctg
     6481 tctatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa
     6541 gagtctatcc tgcccaagag gaacagcgac aagctgatcg ccagaaagaa ggactgggac
     6601 cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc
     6661 aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc
     6721 atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac
     6781 aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa
     6841 aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc
     6901 ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc
     6961 tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaaaca ctacctggac
     7021 gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg
     7081 gacaaggtgc tgagcgccta caacaagcac agagacaagc ctatcagaga gcaggccgag
     7141 aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt
     7201 gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg
     7261 atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc
     7321 gacgcctatc cctatgacgt gcccgattat gccagcctgg gcagcggctc ccccaagaaa
     7381 aaacgcaagg tggaagatcc taagaaaaag cggaaagtgg acggcattgg tagtgggagc
     7441 aacggcagca gcggatccta aaccggtgcg gccgcaatcg atcgcccacc ccaacttgtt
     7501 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     7561 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     7621 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     7681 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     7741 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     7801 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     7861 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     7921 cgttcggctg cggcgagcgg tatcagctca ctcaaaggac ggccgcataa cttcgtataa
     7981 tgtatgctat acgaagttat aagcttgggc tgcaggtcga gggatctcca taagagaaga
     8041 gggacagcta tgactgggag tagtcaggag aggaggaaaa atctggctag taaaacatgt
     8101 aaggaaaatt ttagggatgt taaagaaaaa aataacacaa aacaaaatat aaaaaaaatc
     8161 taacctcaag tcaaggcttt tctatggaat aaggaatgga cagcaggggg ctgtttcata
     8221 tactgatgac ctctttatag ccaacctttg ttcatggcag ccagcatatg ggcatatgtt
     8281 gccaaactct aaaccaaata ctcattctga tgttttaaat gatttgccct cccatatgtc
     8341 cttccgagtg agagacacaa aaaattccaa cacactattg caatgaaaat aaatttcctt
     8401 tattagccag aagtcagatg ctcaaggggc ttcatgatgt ccccataatt tttggcagag
     8461 ggaaaaagat ctcagtggta tttgtgagcc agggcattgg ccacaccagc caccaccttc
     8521 tgataggcag cctgcacctg aggagtgaat tcacgcgttt agcctggcag catatcgaga
     8581 tcgaagtcat cgagagcgtc agcagggagc atatccagat caaaatcatc cagggcatcg
     8641 gcgggcagca tgtccaggtc gaagtcgtcc agggcgtcgg tgggtccgcc gctctcgcac
     8701 ttcagctgct tctcgaggcc gcagatgatc agttccaggc cgaacaggaa ggcgggctcg
     8761 gcgccctgtc tgtcgaacag ctcgatggcc tgcttcagca gggggggcat gctgtcggtg
     8821 gtgggtgtct ctctttcctc tttggcgacc tggtgttcct gttcttccag cacgcagccc
     8881 agggtgaagt ggcccacggc gctcagggcg tacagggcgt tttccaggct gaagccctgc
     8941 tggcacagga aggccagctg gttttccagt gtctcgtact gcttctcggt gggtctggtg
     9001 cccaggtgca ctttggcgcc gtcccggtgg ctcagcaggg cgcatctgta gctcttggcg
     9061 ttgttccgca ggaagtcctg ccagctctcg ccttccagag ggcagctgtg ggtgtggtgc
     9121 cggtccagca tctcgatggg cagggcgtcc agcagggccc gcttgttctt cacgtgccag
     9181 tacagggtgg gctgttccac gcccagtttc tgggccagct tccgggtggt caggccctcg
     9241 atgcccacgc cgttcagcag ttccagggcg ctgttgatca ctttgctctt gtccagccgg
     9301 ctcatgtcga cattctttgc caaaatgatg agacagcaca ataaccagca cgttgcccag
     9361 gagctgtagg aaaaagaaga aggcatgaac atggttagca gaggctctag agccgccggt
     9421 cacacgccag aagccgaacc ccgccctgcc ccgtcccccc cgaaggcagc cgtccccccg
     9481 cggacagccc cgaggctgga gagggagaag gggacggcgg cgcggcgacg cacgaaggcc
     9541 ctccccgccc atttccttcc tgccggcgcc gcaccgcttc gccccgcgcc cgctagaggg
     9601 ggtgcggcgg cgcctcccag atttcggctc cgcacagatt tgggacaaag gaagtccctg
     9661 cgccctctcg cacgattacc ataaaaggca atggctgcgg ctcgccgcgc ctcgacagcc
     9721 gccggcgctc cgggggccgc cgcgcccctc ccccgagccc tccccggccc gaggcggccc
     9781 cgccccgccc ggcaccccca cctgccgcca ccccccgccc ggcacggcga gccccgcgcc
     9841 acgccccgta cggagccccg cacccgaagc cgggccgtgc tcagcaactc ggggaggggg
     9901 gtgcaggggg ggttgcagcc cgaccgacgc gcccacaccc cctgctcacc cccccacgca
     9961 cacaccccgc acgcagcctt tgttcccctc gcagcccccc ccgcaccgcg gggcaccgcc
    10021 cccggccgcg ctcccctcgc gcacactgcg gagcgcacaa agccccgcgc cgcgcccgca
    10081 gcgctcacag ccgccgggca gcgcggagcc gcacgcggcg ctccccacgc acacacacac
    10141 gcacgcaccc cccgagccgc tccccccgca caaagggccc tcccggagcc cctcaaggct
    10201 ttcacgcagc cacagaaaag aaacaagccg tcattaaacc aagcgctaat tacagcccgg
    10261 aggagaaggg ccgtcccgcc cgctcacctg tgggagtaac gcggtcagtc agagccgggg
    10321 cgggcggcgc gaggcggcgg cggagcgggg cacggggcga aggcagcgtc gcagcgactc
    10381 cccgcccgcc gcgcgcttcg ctttttatag ggccgccgcc gccgccgcct cgccataaaa
    10441 ggaaactttc ggagcgcgcc gctctgattg gctgccgccg cacctctccg cctcgccccg
    10501 ccccgcccct cgccccgccc cgccccgcct ggcgcgcgcc cccccccccc ccccgccccc
    10561 atcgctgcac aaaataatta aaaaataaat aaatacaaaa ttgggggtgg ggaggggggg
    10621 gagatgggga gagtgaagca gaacgtgggg ctcacctcga ccatggtaat agcgatgact
    10681 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
    10741 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
    10801 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
    10861 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
    10921 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
    10981 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt
    11041 cgagacccat agagcccacc gcatccccag catgcctgct attgtcttcc caatcctccc
    11101 ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg
    11161 atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg
    11221 aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaggctg
    11281 atcagcgggt ttaaactcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
    11341 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
    11401 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
    11461 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
    11521 catcgccatg ggtcacgacg agatcatcgc cgtcgggcat gcgcgccttg agcctggcga
    11581 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
    11641 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
    11701 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
    11761 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
    11821 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
    11881 ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg
    11941 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
    12001 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
    12061 aacctgcgtg caatccatct tgttcaatgg ccgatcccat ggtggcggta ccctcgagcc
    12121 tagggccggg attctcctcc acgtcaccgc atgttagaag acttcctctg ccctctccgc
    12181 tgccagatct ctcgaggccc tgtgggagga agagaagagg tcagaagctt ataacttcgt
    12241 ataatgtatg ctatacgaag ttattgcccc actgtggggt ggaggggaca gataaaagta
    12301 cccagaacca gagccacatt aaccggccct gggaatataa ggtggtccca gctcggggac
    12361 acaggatccc tggaggcagc aaacatgctg tcctgaagtg gacatagggg cccgggttgg
    12421 aggaagaaga ctagctgagc tctcggaccc ctggaagatg ccatgacagg gggctggaag
    12481 agctagcaca gactagagag gtaagggggg taggggagct gcccaaatga aaggagtgag
    12541 aggtgacccg aatccacagg agaacggggt gtccaggcaa agaaagcaag aggatggaga
    12601 ggtggctaaa gccagggaga cggggtactt tggggttgtc cagaaaaacg gtgatgatgc
    12661 aggcctacaa gaaggggagg cgggacgcaa gggagacatc cgtcggagaa ggccatccta
    12721 agaaacgaga gatggcacag gccccagaag gagaaggaaa agggaaccca gcgagtgaag
    12781 acggcatggg gttgggtgag ggaggagaga tgcccggaga ggacccagac acggggagga
    12841 tccgctcaga ggacatcacg tggtgcagcg ccgagaagga agtgctccgg aaagagcatc
    12901 cttgggcagc aacacagcag agagcaaggg gaagagggag tggaggaaga cggaacctga
    12961 aggaggcggc agggaaggat ctgggccagc cgtagaggtg acccaggcca caagctgcag
    13021 acagaaagcg gcacaggccc aggggagaga atgctggtca gagaaagca
//
