LOCUS       Exported                9553 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses human codon optimized Cas9 D10A mutant which functions as 
            a nickase for genome engineering.
ACCESSION   .
VERSION     .
KEYWORDS    hCas9_D10A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9553)
  AUTHORS   Mali P, Yang L, Esvelt KM, Aach J, Guell M, Dicarlo JE, Norville JE,
            Church GM
  TITLE     RNA-Guided Human Genome Engineering via Cas9.
  JOURNAL   Science. 2013 Jan 3.
  PUBMED    23287722
REFERENCE   2  (bases 1 to 9553)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9553
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      313..322
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             319..4422
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4435..4455
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4510..4529)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4554..4602
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4804..5232
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4891..4910)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5101..5122
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5241..5261)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5246..5575
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5426..5561
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5488..5507
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5642..6436
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5696..5715)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6306..6325
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6612..6733
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6649..6668)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6703..6722
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6782..6798)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6782..6798)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6795..6817)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6806..6822
                     /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(6830..6860)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6875..6896
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7013..7030)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7184..7772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7264..7283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7943..8803)
                     /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     8566..8585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8804..8908)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8983..9002)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        9174..9553
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagag
      301 gatcgaaccc ttgccaccat ggacaagaag tactccattg ggctcgctat cggcacaaac
      361 agcgtcggct gggccgtcat tacggacgag tacaaggtgc cgagcaaaaa attcaaagtt
      421 ctgggcaata ccgatcgcca cagcataaag aagaacctca ttggcgccct cctgttcgac
      481 tccggggaga cggccgaagc cacgcggctc aaaagaacag cacggcgcag atatacccgc
      541 agaaagaatc ggatctgcta cctgcaggag atctttagta atgagatggc taaggtggat
      601 gactctttct tccataggct ggaggagtcc tttttggtgg aggaggataa aaagcacgag
      661 cgccacccaa tctttggcaa tatcgtggac gaggtggcgt accatgaaaa gtacccaacc
      721 atatatcatc tgaggaagaa gcttgtagac agtactgata aggctgactt gcggttgatc
      781 tatctcgcgc tggcgcatat gatcaaattt cggggacact tcctcatcga gggggacctg
      841 aacccagaca acagcgatgt cgacaaactc tttatccaac tggttcagac ttacaatcag
      901 cttttcgaag agaacccgat caacgcatcc ggagttgacg ccaaagcaat cctgagcgct
      961 aggctgtcca aatcccggcg gctcgaaaac ctcatcgcac agctccctgg ggagaagaag
     1021 aacggcctgt ttggtaatct tatcgccctg tcactcgggc tgacccccaa ctttaaatct
     1081 aacttcgacc tggccgaaga tgccaagctt caactgagca aagacaccta cgatgatgat
     1141 ctcgacaatc tgctggccca gatcggcgac cagtacgcag accttttttt ggcggcaaag
     1201 aacctgtcag acgccattct gctgagtgat attctgcgag tgaacacgga gatcaccaaa
     1261 gctccgctga gcgctagtat gatcaagcgc tatgatgagc accaccaaga cttgactttg
     1321 ctgaaggccc ttgtcagaca gcaactgcct gagaagtaca aggaaatttt cttcgatcag
     1381 tctaaaaatg gctacgccgg atacattgac ggcggagcaa gccaggagga attttacaaa
     1441 tttattaagc ccatcttgga aaaaatggac ggcaccgagg agctgctggt aaagcttaac
     1501 agagaagatc tgttgcgcaa acagcgcact ttcgacaatg gaagcatccc ccaccagatt
     1561 cacctgggcg aactgcacgc tatcctcagg cggcaagagg atttctaccc ctttttgaaa
     1621 gataacaggg aaaagattga gaaaatcctc acatttcgga taccctacta tgtaggcccc
     1681 ctcgcccggg gaaattccag attcgcgtgg atgactcgca aatcagaaga gaccatcact
     1741 ccctggaact tcgaggaagt cgtggataag ggggcctctg cccagtcctt catcgaaagg
     1801 atgactaact ttgataaaaa tctgcctaac gaaaaggtgc ttcctaaaca ctctctgctg
     1861 tacgagtact tcacagttta taacgagctc accaaggtca aatacgtcac agaagggatg
     1921 agaaagccag cattcctgtc tggagagcag aagaaagcta tcgtggacct cctcttcaag
     1981 acgaaccgga aagttaccgt gaaacagctc aaagaagact atttcaaaaa gattgaatgt
     2041 ttcgactctg ttgaaatcag cggagtggag gatcgcttca acgcatccct gggaacgtat
     2101 cacgatctcc tgaaaatcat taaagacaag gacttcctgg acaatgagga gaacgaggac
     2161 attcttgagg acattgtcct cacccttacg ttgtttgaag atagggagat gattgaagaa
     2221 cgcttgaaaa cttacgctca tctcttcgac gacaaagtca tgaaacagct caagaggcgc
     2281 cgatatacag gatgggggcg gctgtcaaga aaactgatca atgggatccg agacaagcag
     2341 agtggaaaga caatcctgga ttttcttaag tccgatggat ttgccaacag gaacttcatg
     2401 cagttgatcc atgatgactc tctcaccttt aaggaggaca tccagaaagc acaagtttct
     2461 ggccaggggg acagtcttca cgagcacatc gctaatcttg caggtagccc agctatcaaa
     2521 aagggaatac tgcagaccgt taaggtcgtg gatgaactcg tcaaagtaat gggaaggcat
     2581 aagcccgaga atatcgttat cgagatggcc cgagagaacc aaactaccca gaagggacag
     2641 aagaacagta gggaaaggat gaagaggatt gaagagggta taaaagaact ggggtcccaa
     2701 atccttaagg aacacccagt tgaaaacacc cagcttcaga atgagaagct ctacctgtac
     2761 tacctgcaga acggcaggga catgtacgtg gatcaggaac tggacatcaa tcggctctcc
     2821 gactacgacg tggatcatat cgtgccccag tcttttctca aagatgattc tattgataat
     2881 aaagtgttga caagatccga taaaaataga gggaagagtg ataacgtccc ctcagaagaa
     2941 gttgtcaaga aaatgaaaaa ttattggcgg cagctgctga acgccaaact gatcacacaa
     3001 cggaagttcg ataatctgac taaggctgaa cgaggtggcc tgtctgagtt ggataaagcc
     3061 ggcttcatca aaaggcagct tgttgagaca cgccagatca ccaagcacgt ggcccaaatt
     3121 ctcgattcac gcatgaacac caagtacgat gaaaatgaca aactgattcg agaggtgaaa
     3181 gttattactc tgaagtctaa gctggtctca gatttcagaa aggactttca gttttataag
     3241 gtgagagaga tcaacaatta ccaccatgcg catgatgcct acctgaatgc agtggtaggc
     3301 actgcactta tcaaaaaata tcccaagctt gaatctgaat ttgtttacgg agactataaa
     3361 gtgtacgatg ttaggaaaat gatcgcaaag tctgagcagg aaataggcaa ggccaccgct
     3421 aagtacttct tttacagcaa tattatgaat tttttcaaga ccgagattac actggccaat
     3481 ggagagattc ggaagcgacc acttatcgaa acaaacggag aaacaggaga aatcgtgtgg
     3541 gacaagggta gggatttcgc gacagtccgg aaggtcctgt ccatgccgca ggtgaacatc
     3601 gttaaaaaga ccgaagtaca gaccggaggc ttctccaagg aaagtatcct cccgaaaagg
     3661 aacagcgaca agctgatcgc acgcaaaaaa gattgggacc ccaagaaata cggcggattc
     3721 gattctccta cagtcgctta cagtgtactg gttgtggcca aagtggagaa agggaagtct
     3781 aaaaaactca aaagcgtcaa ggaactgctg ggcatcacaa tcatggagcg atcaagcttc
     3841 gaaaaaaacc ccatcgactt tctcgaggcg aaaggatata aagaggtcaa aaaagacctc
     3901 atcattaagc ttcccaagta ctctctcttt gagcttgaaa acggccggaa acgaatgctc
     3961 gctagtgcgg gcgagctgca gaaaggtaac gagctggcac tgccctctaa atacgttaat
     4021 ttcttgtatc tggccagcca ctatgaaaag ctcaaagggt ctcccgaaga taatgagcag
     4081 aagcagctgt tcgtggaaca acacaaacac taccttgatg agatcatcga gcaaataagc
     4141 gaattctcca aaagagtgat cctcgccgac gctaacctcg ataaggtgct ttctgcttac
     4201 aataagcaca gggataagcc catcagggag caggcagaaa acattatcca cttgtttact
     4261 ctgaccaact tgggcgcgcc tgcagccttc aagtacttcg acaccaccat agacagaaag
     4321 cggtacacct ctacaaagga ggtcctggac gccacactga ttcatcagtc aattacgggg
     4381 ctctatgaaa caagaatcga cctctctcag ctcggtggag acagcagggc tgaccccaag
     4441 aagaagagga aggtgtgaaa gggttcgatc cctaccggtt agtaatgagt ttaaacgggg
     4501 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     4561 aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     4621 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     4681 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     4741 acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc
     4801 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4861 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4921 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4981 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     5041 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5101 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     5161 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     5221 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     5281 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     5341 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5401 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5461 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     5521 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     5581 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     5641 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5701 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5761 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5821 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5881 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5941 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     6001 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     6061 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     6121 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     6181 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     6241 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     6301 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     6361 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     6421 tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca
     6481 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     6541 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6601 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6661 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6721 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     6781 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6841 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6901 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6961 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     7021 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     7081 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     7141 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7201 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7261 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7321 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     7381 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7441 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7501 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7561 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7621 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7681 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7741 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7801 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7861 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7921 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7981 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     8041 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     8101 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     8161 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     8221 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     8281 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     8341 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8401 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8461 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8521 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8581 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8641 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8701 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8761 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8821 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8881 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     8941 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     9001 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     9061 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     9121 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
     9181 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     9241 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     9301 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     9361 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     9421 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     9481 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     9541 cgctattacc atg
//
