LOCUS       miniCGBE1-SpRY-Cas9_(pBM1571)        9195 bp    DNA     circular SYN 06-APR-2021
DEFINITION  pCMV-rAPOBEC1(R33A)-SpRY-nCas9(D10A)-P2A-EGFP.
ACCESSION   urn.local...3bk-d7uhdgy
KEYWORDS    .
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     CDS             6..23
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /translation="HHHHHH"
                     /label="6xHis"
     primer_bind     complement(46..63)
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
                     /label="BGH-rev"
     polyA_signal    52..276
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     primer_bind     complement(347..363)
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /label="M13 rev"
     primer_bind     complement(347..363)
                     /note="In lacZ gene. Also called M13-rev"
                     /label="M13 Reverse"
     primer_bind     complement(360..382)
                     /note="In lacZ gene"
                     /label="M13/pUC Reverse"
     protein_bind    371..387
                     /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)."
                     /label="lac operator"
     promoter        complement(395..425)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    440..461
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                      of cAMP."
                     /label="CAP binding site"
     primer_bind     complement(578..595)
                     /note="L4440 vector, forward primer"
                     /label="L4440"
     rep_origin      complement(749..1337)
                     /direction="LEFT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /label="ori"
     primer_bind     complement(829..848)
                     /note="pBR322 origin, forward primer"
                     /label="pBR322ori-F"
     CDS             complement(1508..2368)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /label="AmpR"
     primer_bind     2131..2150
                     /note="Ampicillin resistance gene, reverse primer"
                     /label="Amp-R"
     promoter        complement(2369..2473)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     complement(2552..2571)
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
                     /label="pRS-marker"
     enhancer        2743..3122
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        3123..3326
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /label="CMV promoter"
     primer_bind     3277..3297
                     /note="Human CMV immediate early promoter, forward primer"
                     /label="CMV-F"
     misc_feature    3360..3367
                     /Recognition_pattern="GC^GGCCGC"
                     /annotation_group="GCGGCCGC"
                     /label="NotI"
                     /note="Geneious type: restriction site"
     promoter        3368..3386
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     motif           3391..>3402
                     /created_by="User"
                     /modified_by="User"
                     /label="KOZAC"
     ORF             3400..9195
                     /Genetic_code="Standard"
                     /Reading_frame=1
                     /created_by="ORF Finder"
                     /label="ORF 1 (frame 1)"
     primer_bind     3403..3456
                     /created_by="User"
                     /Sequence="AAACGGACAGCCGACGGAAGCGAGTTCGAGTCACCAAAGAAGAAGCG
                     GAAAGTC"
                     /Color="#0077ee"
                     /modified_by="User"
                     /label="bpNLS"
     CDS             3457..4140
                     /created_by="jonlevy"
                     /Color="#ff9911"
                     /modified_by="User"
                     /label="Genscript rAPOBEC1 (codon-optimized)"
     mutation        3550..3552
                     /created_by="User"
                     /label="R33A"
     CDS             4141..4236
                     /created_by="jonlevy"
                     /Transferred_From="JL545-pCMV_BPNLS_rAPOBEC1(genscript)_dl
                     BE4_NpuN1"
                     /Transferred_Similarity="100.00%"
                     /Color="#666666"
                     /modified_by="User"
                     /label="Genscript BE4 linker"
     CDS             <4237..8337
                     /product="nickase mutant of the Cas9 endonuclease from the
                      Streptococcus pyogenes Type II CRISPR/Cas system"
                     /note="generates RNA-guided single strand nicks in DNA due
                      to the D10A mutation in the RuvC catalytic domain"
                     /Original_Translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLG
                     NTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVD
                     DSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLR
                     LIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKA
                     ILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSK
                     DTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYD
                     EHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMD
                     GTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEK
                     ILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDK
                     NLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRK
                     VTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDIL
                     EDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQ
                     SGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPA
                     IKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKE
                     LGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLK
                     DDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERG
                     GLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVS
                     DFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMI
                     AKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDF
                     ATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPT
                     VAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLII
                     KLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQ
                     KQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHL
                     FTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"
                     /modified_by="Kleinstiver Lab 1"
                     /label="nSpCas9(D10A)-RRLQKRRPQR"
     misc_difference 4261..4263
                     /created_by="bpk6"
                     /label="D10A"
     misc_difference 4414..4416
                     /created_by="Kleinstiver Lab 1"
                     /label="A61R"
     misc_difference 7564..7566
                     /created_by="Kleinstiver Lab 1"
                     /label="L1111R"
     misc_difference 7636..7641
                     /created_by="Kleinstiver Lab 1"
                     /label="D1135L/S1136W"
     misc_difference 7885..7890
                     /created_by="Kleinstiver Lab 1"
                     /label="G1218K/E1219Q"
     misc_difference 8182..8184
                     /created_by="Kleinstiver Lab 1"
                     /label="N1317R"
     misc_difference 8197..8199
                     /created_by="Kleinstiver Lab 1"
                     /label="A1322R"
     misc_difference 8230..8232
                     /created_by="Kleinstiver Lab 1"
                     /label="R1333P"
     misc_difference 8236..8238
                     /created_by="Kleinstiver Lab 1"
                     /label="R1335Q"
     misc_difference 8242..8244
                     /created_by="Kleinstiver Lab 1"
                     /label="T1337R"
     primer_bind     8338..8349
                     /Sequence="TCTGGCGGCTCA"
                     /Hairpin_Tm="None"
                     /Tm="47.1"
                     /Self_Dimer_Tm="11.4"
                     /%GC="66.7"
                     /created_by="primer3"
                     /Color="#666666"
                     /modified_by="User"
                     /label="SGGS"
     misc_feature    8350..>8379
                     /created_by="User"
                     /Color="#1199ff"
                     /label="SV40 NLS"
     misc_feature    8380..8400
                     /created_by="User"
                     /Color="#1199ff"
                     /label="SV40 NLS"
     CDS             8401..8409
                     /created_by="User"
                     /Color="#ff7400"
                     /modified_by="User"
                     /label="linker"
     CDS             8410..8475
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIK
                     KNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLE
                     ESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAH
                     MIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSK
                     SRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLD
                     NLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTL
                     LKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVK
                     LNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPY
                     YVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVL
                     PKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKE
                     DYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLT
                     LFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDF
                     LKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQT
                     VKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKE
                     HPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKV
                     LTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKA
                     GFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF
                     YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIG
                     KATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS
                     MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVV
                     AKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLF
                     ELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQH
                     KHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGA
                     PAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDKRPAATKKAG
                     QAKKKKDYKDDDDKGSGATNFSLLKQAGDVEENPGPTEYKPTVRLATRDDVPRAVRTL
                     AAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPES
                     VEAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGL
                     GSAVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKP
                     GA*"
                     /Color="#5500cc"
                     /modified_by="User"
                     /label="P2A"
     CDS             8476..9192
                     /created_by="User"
                     /Color="#55ff55"
                     /modified_by="User"
                     /label="eGFP"
     primer_bind     9193..9195
                     /Sequence="TGA"
                     /Color="#000000"
                     /created_by="User"
                     /label="STOP"
     misc_feature    9195..5
                     /Recognition_pattern="A^CCGGT"
                     /annotation_group="ACCGGT"
                     /label="AgeI"
                     /note="Geneious type: restriction site"
ORIGIN      
        1 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt
       61 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
      121 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
      181 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
      241 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
      301 ggggctcgat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
      361 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
      421 gtaaagccta gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
      481 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
      541 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
      601 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      661 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      721 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
      781 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
      841 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
      901 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
      961 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1021 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1081 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1141 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1201 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1261 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1321 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac actcagtgga
     1381 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1441 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1501 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1561 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     1621 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     1681 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     1741 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     1801 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     1861 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     1921 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     1981 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2041 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     2101 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     2161 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     2221 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     2281 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     2341 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2401 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2461 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     2521 gatctcccga tcccctaggg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
     2581 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     2641 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     2701 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
     2761 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     2821 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     2881 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     2941 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     3001 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     3061 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     3121 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     3181 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     3241 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     3301 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagagatccg
     3361 cggccgctaa tacgactcac tatagggaga gccgccacca tgaaacggac agccgacgga
     3421 agcgagttcg agtcaccaaa gaagaagcgg aaagtctcct cagagactgg gcctgtcgcc
     3481 gtcgatccaa ccctgcgccg ccggattgaa cctcacgagt ttgaagtgtt ctttgacccc
     3541 cgggagctgg ccaaggagac atgcctgctg tacgagatca actggggagg caggcactcc
     3601 atctggaggc acacctctca gaacacaaat aagcacgtgg aggtgaactt catcgagaag
     3661 tttaccacag agcggtactt ctgccccaat accagatgta gcatcacatg gtttctgagc
     3721 tggtcccctt gcggagagtg tagcagggcc atcaccgagt tcctgtccag atatccacac
     3781 gtgacactgt ttatctacat cgccaggctg tatcaccacg cagacccaag gaataggcag
     3841 ggcctgcgcg atctgatcag ctccggcgtg accatccaga tcatgacaga gcaggagtcc
     3901 ggctactgct ggcggaactt cgtgaattat tctcctagca acgaggccca ctggcctagg
     3961 tacccacacc tgtgggtgcg cctgtacgtg ctggagctgt attgcatcat cctgggcctg
     4021 cccccttgtc tgaatatcct gcggagaaag cagccccagc tgaccttctt tacaatcgcc
     4081 ctgcagtctt gtcactatca gaggctgcca ccccacatcc tgtgggccac aggcctgaag
     4141 tctggaggat ctagcggagg atcctctggc agcgagacac caggaacaag cgagtcagca
     4201 acaccagaga gcagtggcgg cagcagcggc ggcagcgaca agaagtacag catcggcctg
     4261 gccatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc
     4321 aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga
     4381 gccctgctgt tcgacagcgg cgaaacagcc gagagaaccc ggctgaagag aaccgccaga
     4441 agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag
     4501 atggccaagg tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag
     4561 gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac
     4621 gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc
     4681 gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg
     4741 atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg
     4801 cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag
     4861 gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg
     4921 cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc
     4981 cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac
     5041 acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg
     5101 tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac
     5161 accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac
     5221 caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag
     5281 attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag
     5341 gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg
     5401 ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc
     5461 atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt
     5521 tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc
     5581 tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc
     5641 gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag
     5701 agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc
     5761 aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac
     5821 gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg
     5881 gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc
     5941 aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc
     6001 tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat
     6061 gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga
     6121 gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag
     6181 cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc
     6241 atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc
     6301 aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag
     6361 aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc
     6421 agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa
     6481 gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc
     6541 acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa
     6601 gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag
     6661 aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac
     6721 atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctgaaggac
     6781 gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac
     6841 gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc
     6901 aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc
     6961 gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag
     7021 cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg
     7081 atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat
     7141 ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg
     7201 aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg
     7261 tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc
     7321 ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag
     7381 attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc
     7441 ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg
     7501 ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct
     7561 atcagaccca agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag
     7621 aagtacggcg gcttcctgtg gcccaccgtg gcctattctg tgctggtggt ggccaaagtg
     7681 gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg
     7741 gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa
     7801 gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc
     7861 cggaagagaa tgctggcctc tgccaagcag ctgcagaagg gaaacgaact ggccctgccc
     7921 tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc
     7981 gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc
     8041 atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa
     8101 gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc
     8161 atccacctgt ttaccctgac cagactggga gcccctagag ccttcaagta ctttgacacc
     8221 accatcgacc ccaagcagta cagaagcacc aaagaggtgc tggacgccac cctgatccac
     8281 cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggtgactct
     8341 ggcggctcaa aaagaaccgc cgacggcagc gaattcgagc ccaagaagaa gaggaaagtc
     8401 ggaggcggtg gaagcggcgc aacaaacttc tctctgctga aacaagccgg agatgtcgaa
     8461 gagaatcctg gaccgatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     8521 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     8581 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     8641 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     8701 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     8761 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     8821 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     8881 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     8941 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     9001 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     9061 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     9121 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     9181 gagctgtaca agtga
//
