LOCUS       Exported                9797 bp ds-DNA     circular SYN 18-FEB-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9797)
  AUTHORS   Institute of Molecular and Genomic Medicine
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Feb 18, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9797
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     2..22
                     /label=T3
                     /note="T3 promoter, forward primer"
     primer_bind     5..24
                     /label=mission_T3 promoter seq
     primer_bind     complement(22..42)
                     /label=Primer 2
     primer_bind     complement(23..48)
                     /label=pp438-EcoRI-longum core-F
     primer_bind     complement(25..48)
                     /label=Primer 3
     primer_bind     42..68
                     /label=pp455-EcoRI-gapdh-F
     primer_bind     43..69
                     /label=pp137
     promoter        49..333
                     /label=P3
     primer_bind     314..352
                     /label=pp458-gapdh-Cas9-F
     primer_bind     complement(314..343)
                     /label=pp482-BbsI-gapdh-R
     primer_bind     complement(314..336)
                     /label=Primer 1a
     primer_bind     complement(314..333)
                     /label=pp456-gapdh-R
     CDS             334..1020
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     primer_bind     334..353
                     /label=pp457-kasOp-Cas9-F
     primer_bind     334..352
                     /label=APOBEC-F
     primer_bind     335..354
                     /label=pp483-BbsI-Cas9-F
     primer_bind     complement(475..493)
                     /label=pp494-APO_EcoRI site
     CDS             1069..5169
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     1260..1270
                     /label=Primer 1
     primer_bind     1290..1311
                     /label=Primer 4
     primer_bind     complement(1314..1337)
                     /label=Cas9-R
     CDS             5182..5433
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     terminator      5614..5662
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     primer_bind     complement(5741..5768)
                     /label=pp459-Cas9-R
     primer_bind     5763..5842
                     /label=pp452-SpeI-tcp830-F
     primer_bind     5763..5790
                     /label=pp442-SpeI-tcp830-sgRNA-F
     promoter        5769..5821
                     /label=tcp830
                     /note="substantial leakiness based on siegl 2013"
     misc_feature    5822..6410
                     /label=golden gate compatible sgRNA insertion cassette
     primer_bind     5822..5842
                     /label=Primer 1
     primer_bind     5822..5842
                     /label=pp451-SpeI-kasOP17tss-F
     misc_feature    5824..5829
                     /label=BbsI
     CDS             complement(5830..6126)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSG"
     primer_bind     5976..5998
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5990..6007
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6017..6036
                     /label=T7
                     /note="T7 promoter, forward primer"
     primer_bind     6068..6083
                     /label=Primer 2
     primer_bind     6068..6082
                     /label=pp438-EcoRI-longum core-F
     primer_bind     6068..6080
                     /label=Primer 3
     primer_bind     complement(6081..6100)
                     /label=mission_T3 promoter seq
     primer_bind     complement(6083..6103)
                     /label=T3
                     /note="T3 promoter, forward primer"
     primer_bind     complement(6122..6143)
                     /label=mission_M13R
     primer_bind     complement(6122..6138)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6135..6157)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6146..6162
                     /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(6170..6200)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6215..6236
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    6271..6276
                     /label=BbsI
     misc_RNA        6279..6354
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      6376..6410
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     rep_origin      complement(6452..7040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6532..6551)
                     /label=pp235-pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(7018..7040)
                     /label=Primer 2
     primer_bind     complement(7019..7049)
                     /label=pp440-NotI-ori-sgRNA-R
     primer_bind     7039..7069
                     /label=pp439-NotI-longum core-R
     misc_feature    7049..9797
                     /label=pMDY23 backbone
     promoter        7049..7251
                     /label=Pcat
     primer_bind     7049..7075
                     /label=pp123_IM206 primer
     CDS             7249..7908
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=chloramphenicol resistance
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQNCQIGLMTGFYNMR"
     primer_bind     complement(7274..7293)
                     /label=pp495_CAM-NotI site
     primer_bind     complement(7949..8003)
                     /label=pp437-mut NseI to NsiI-F
     primer_bind     complement(8008..8038)
                     /label=pp436-mut BbsI-F
     CDS             complement(8323..8640)
                     /codon_start=1
                     /transl_table=11
                     /gene="repB"
                     /product="RepB"
                     /label=repB
                     /note="pNCC293 replication protein"
                     /protein_id="ABI18631.1"
                     /translation="MVRTTLRKKRPVSARELAEAYGVSTRTIQSWVAMKREDWIDEQAA
                     MREAVRSYHDDEGHTWPQTAEHFNMSQGAVRQRCYRARKEREDEAAEKSKHLPGEIPLF
                     D"
     gene            complement(8323..8640)
                     /gene="repB"
                     /label=repB
     CDS             complement(8634..9554)
                     /codon_start=1
                     /transl_table=11
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="pNCC293 replication protein"
                     /protein_id="ABI18630.1"
                     /translation="MYAVAMSDEYSQPTLELSRTFEGWWLPERPLCCDDDYSQLHRRSR
                     ADALKCKHIEANPAALVNTIVVDIDDANAKAMALWEHEGMRPNWIAENPANGHAHAGWV
                     LTFPVPRTDLARLKPLKLLHATTEGLRRSCDGDMGYSGLLMKNPEHPAWASDIIEWDTY
                     DLEQLVQSLQEHGDMPPVSWKRTKRARTQGLGRNCTLFDKARTLAYRYVRRLPDRSEAS
                     SEALRLYVRRTCHELNVSLFPDPLHAREVEDIAKSIHKWIVTRSRMWLDGAIANAATFI
                     AIQSARGHKHGENKYQQVMKEALEW"
     gene            complement(8634..9554)
                     /gene="repA"
                     /label=repA
     primer_bind     9700..9719
                     /label=pp154
     primer_bind     complement(9777..9797)
                     /label=Primer 1
ORIGIN
        1 tgcaattaac cctcactaaa gggaacaaaa gctggagctc cagaattcgc tgctccttcg
       61 gtcggacgtg cgtctacggg caccttaccg cagccgtcgg ctgtgcgaca cggacggatc
      121 gggcgaactg gccgatgctg ggagaagcgc gctgctgtac ggcgcgcacc gggtgcggag
      181 cccctcggcg agcggtgtga aacttctgtg aatggcctgt tcggttgctt tttttatacg
      241 gctgccagat aaggcttgca gcatctgggc ggctaccgct atgatcgggg cgttcctgca
      301 attcttagtg cgagtatctg aaaggggata cgcatgtcct ccgagaccgg ccccgtggcc
      361 gtggacccga ccctgcgtcg ccgcatcgag ccgcacgagt tcgaggtgtt cttcgacccg
      421 cgcgagctgc gcaaggagac ctgcctgctg tacgagatca actggggcgg ccggcactcc
      481 atctggcgcc acacctcgca gaacaccaac aagcacgtcg aggtcaactt catcgagaag
      541 ttcaccaccg agcggtactt ctgcccgaac acccgctgct ccatcacctg gttcctgtcc
      601 tggtccccct gcggcgagtg ctcccgggcc atcaccgagt tcctgtcccg ctacccgcac
      661 gtcaccctgt tcatctacat cgcccggctg taccaccacg ccgacccgcg caaccggcag
      721 ggcctgcgcg acctgatctc ctccggcgtc accatccaga tcatgaccga gcaggagtcc
      781 ggctactgct ggcggaactt cgtcaactac tccccgtcca acgaggccca ctggccccgc
      841 tacccgcacc tgtgggtccg cctgtacgtc ctggagctgt actgcatcat cctgggcctg
      901 ccgccctgcc tgaacatcct gcgtcgcaag cagccgcagc tgaccttctt caccatcgcc
      961 ctccagtcct gccactacca gcgcctgccg ccccacatcc tgtgggccac cggcctgaag
     1021 tccggctccg agaccccggg cacctcggag tccgccaccc cggagtccga caagaagtac
     1081 tccatcggcc tcgcgatcgg caccaactcc gtgggctggg cggtcatcac cgacgagtac
     1141 aaggtcccct ccaagaagtt caaggtcctg ggcaacaccg accggcactc gatcaagaag
     1201 aacctgatcg gcgccctgct cttcgacagc ggcgagaccg ccgaggcgac ccgcctgaag
     1261 cggaccgcgc gtcgccgcta cacccggcgc aagaaccgca tctgctacct gcaggaaatc
     1321 ttctccaacg agatggccaa ggtggacgac tcgttcttcc accgcctgga ggagagcttc
     1381 ctggtggagg aggacaagaa gcacgagcgc cacccgatct tcggcaacat cgtggacgag
     1441 gtggcctacc acgagaagta ccccaccatc taccacctcc gcaagaagct ggtggactcg
     1501 accgacaagg cggacctgcg gctcatctac ctggccctcg cgcacatgat caagttccgc
     1561 ggccacttcc tcatcgaggg cgacctgaac ccggacaact ccgacgtgga caagctcttc
     1621 atccagctgg tgcagaccta caaccagctg ttcgaggaga accccatcaa cgccagcggc
     1681 gtggacgcca aggcgatcct ctccgcgcgc ctgagcaagt cccggcgcct ggagaacctc
     1741 atcgcccagc tgccgggcga gaagaagaac ggcctcttcg gcaacctgat cgcgctgtcg
     1801 ctcggcctga cccccaactt caagagcaac ttcgacctgg ccgaggacgc gaagctccag
     1861 ctgtccaagg acacctacga cgacgacctg gacaacctgc tcgcccagat cggcgaccag
     1921 tacgcggacc tcttcctggc cgcgaagaac ctctcggacg ccatcctgct cagcgacatc
     1981 ctgcgggtca acaccgagat caccaaggcc ccgctgtcgg cgagcatgat caagcggtac
     2041 gacgagcacc accaggacct gaccctgctc aaggccctcg tgcgccagca gctgcccgag
     2101 aagtacaagg aaatcttctt cgaccagtcc aagaacggct acgccggcta catcgacggc
     2161 ggcgcgtcgc aggaggagtt ctacaagttc atcaagccga tcctggagaa gatggacggc
     2221 accgaggagc tgctcgtcaa gctgaaccgc gaggacctgc tccgcaagca gcggaccttc
     2281 gacaacggct ccatcccgca ccagatccac ctgggcgagc tccacgccat cctccggcgc
     2341 caggaggact tctacccctt cctgaaggac aaccgcgaga agatcgagaa gatcctgacc
     2401 ttccgcatcc cgtactacgt cggccccctg gcccgcggca actcccggtt cgcgtggatg
     2461 acccggaagt cggaggagac catcaccccg tggaacttcg aggaggtcgt ggacaagggc
     2521 gcgtccgcgc agtcgttcat cgagcgcatg accaacttcg acaagaacct cccgaacgag
     2581 aaggtcctgc ccaagcactc cctgctctac gagtacttca ccgtgtacaa cgagctgacc
     2641 aaggtcaagt acgtgaccga gggcatgcgg aagccggcct tcctgtcggg cgagcagaag
     2701 aaggcgatcg tggacctgct cttcaagacc aaccgcaagg tcaccgtgaa gcagctgaag
     2761 gaggactact tcaagaagat cgagtgcttc gactccgtcg agatcagcgg cgtggaggac
     2821 cgcttcaacg cctccctggg cacctaccac gacctgctca agatcatcaa ggacaaggac
     2881 ttcctcgaca acgaggagaa cgaggacatc ctggaggaca tcgtcctcac cctgaccctc
     2941 ttcgaggacc gcgagatgat cgaggagcgg ctcaagacct acgcccacct gttcgacgac
     3001 aaggtgatga agcagctgaa gcgtcgccgc tacaccggct ggggccgcct ctcccggaag
     3061 ctgatcaacg gcatccggga caagcagagc ggcaagacca tcctggactt cctcaagtcc
     3121 gacggcttcg ccaaccgcaa cttcatgcag ctcatccacg acgacagcct gaccttcaag
     3181 gaggacatcc agaaggccca ggtctcgggc cagggcgaca gcctccacga gcacatcgcc
     3241 aacctggcgg gctccccggc gatcaagaag ggcatcctcc agaccgtcaa ggtcgtggac
     3301 gagctggtca aggtgatggg ccgccacaag cccgagaaca tcgtgatcga gatggcccgg
     3361 gagaaccaga ccacccagaa gggccagaag aactcgcgcg agcggatgaa gcggatcgag
     3421 gagggcatca aggagctcgg cagccagatc ctgaaggagc acccggtcga gaacacccag
     3481 ctgcagaacg agaagctgta cctctactac ctgcagaacg gccgcgacat gtacgtggac
     3541 caggagctcg acatcaaccg gctgtccgac tacgacgtgg accacatcgt gccgcagtcc
     3601 ttcctgaagg acgactcgat cgacaacaag gtcctgaccc gctcggacaa gaaccggggc
     3661 aagtccgaca acgtgccctc ggaggaggtc gtgaagaaga tgaagaacta ctggcgccag
     3721 ctgctcaacg ccaagctcat cacccagcgc aagttcgaca acctgaccaa ggccgagcgg
     3781 ggcggcctga gcgagctcga caaggcgggc ttcatcaagc gccagctggt cgagacccgg
     3841 cagatcacca agcacgtggc ccagatcctg gactcccgga tgaacaccaa gtacgacgag
     3901 aacgacaagc tgatccgcga ggtcaaggtg atcaccctca agagcaagct ggtctccgac
     3961 ttccgcaagg acttccagtt ctacaaggtc cgggagatca acaactacca ccacgcccac
     4021 gacgcgtacc tgaacgccgt cgtgggcacc gcgctgatca agaagtaccc gaagctggag
     4081 tccgagttcg tctacggcga ctacaaggtc tacgacgtgc gcaagatgat cgccaagagc
     4141 gagcaggaga tcggcaaggc caccgcgaag tacttcttct actccaacat catgaacttc
     4201 ttcaagaccg agatcaccct ggccaacggc gagatccgca agcggcccct gatcgagacc
     4261 aacggcgaga ccggcgagat cgtctgggac aagggccgcg acttcgccac cgtccggaag
     4321 gtgctgtcga tgccgcaggt caacatcgtg aagaaaaccg aggtgcagac cggcggcttc
     4381 agcaaggagt ccatcctccc caagcgcaac agcgacaagc tgatcgcccg gaagaaggac
     4441 tgggacccga agaagtacgg cggcttcgac agccccaccg tcgcctactc cgtgctggtc
     4501 gtggcgaagg tcgagaaggg caagagcaag aagctgaagt ccgtgaagga gctgctcggc
     4561 atcaccatca tggagcgctc ctcgttcgag aagaacccga tcgacttcct ggaggccaag
     4621 ggctacaagg aggtcaagaa ggacctcatc atcaagctgc ccaagtacag cctgttcgag
     4681 ctggagaacg gccgcaagcg gatgctcgcc tccgcgggcg agctgcagaa gggcaacgag
     4741 ctggccctcc cgtcgaagta cgtcaacttc ctgtacctcg cgtcccacta cgagaagctg
     4801 aagggctcgc ccgaggacaa cgagcagaag cagctcttcg tggagcagca caagcactac
     4861 ctggacgaga tcatcgagca gatcagcgag ttcagcaagc gcgtcatcct ggccgacgcg
     4921 aacctcgaca aggtgctgtc cgcctacaac aagcaccgcg acaagccgat ccgggagcag
     4981 gcggagaaca tcatccacct gttcaccctc accaacctgg gcgcccccgc cgcgttcaag
     5041 tacttcgaca ccaccatcga ccgcaagcgg tacacctcca ccaaggaggt cctcgacgcg
     5101 accctgatcc accagagcat caccggcctg tacgagaccc gcatcgacct gtcccagctc
     5161 ggcggcgact ccggcggctc caccaacctg tccgacatca tcgagaagga gaccggcaag
     5221 cagctggtca tccaggagtc catcctgatg ctgccggagg aagtcgagga agtcatcggc
     5281 aacaagcccg agtccgacat cctggtccac accgcctacg acgagtccac cgacgagaac
     5341 gtcatgctgc tgacctcgga cgcccccgag tacaagccct gggccctggt catccaggac
     5401 tccaacggcg agaacaagat caagatgctg tgatctagag gatccccggg taccgagctc
     5461 ggattcccca gatctaaagt tttgtcgtct ttccagacgt tagtaaatga attttctgta
     5521 tgaggttttg ctaaacaact ttcaacagtt tcagcggagt gagaatagaa aggaacaact
     5581 aaaggaattg cgaataataa ttttttcacg ttgaaaatct ccaaaaaaaa aggctccaaa
     5641 aggagccttt aattgtatcg gtttatcagc ttgctttcga ggtgaatttc ttaaacagct
     5701 tgataccgat agttgcgccg acaatgacaa caaccatcgc ccacgcataa ccgatatatt
     5761 cgactagtct gttggctact ctatcattga tagtggtagg atccctatca gtgatagaga
     5821 taagtcttct cagccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
     5881 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5941 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     6001 ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct
     6061 cgaggtcctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     6121 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6181 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6241 cgttgcgctc actgcccgct ttccaccggt gaagacatgt tttagagcta gaaatagcaa
     6301 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgcttttta
     6361 ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat ctagaggcca
     6421 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6481 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6541 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6601 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6661 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gacccccccg
     6721 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6781 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6841 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6901 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6961 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7021 gcagaaaaaa aggatctcaa gcggccgcat cccattatgc tttggcagtt tattcttgac
     7081 atgtagtgag ggggctggta taatcacata cggccgataa agcaagcata taatattgcg
     7141 tttcatcttt agaagcgaat ttcgccaata ttataattat caaagagagg ggtggcaaac
     7201 ggtatttggc attattaggt taaaaaatgt agaaggagag tgaaacccat gaactttaat
     7261 aaaattgatt tagacaattg gaagagaaaa gagatattta atcattattt gaaccaacaa
     7321 acgactttta gtataaccac agaaattgat attagtgttt tataccgaaa cataaaacaa
     7381 gaaggatata aattttaccc tgcatttatt ttcttagtga caagggtgat aaactcaaat
     7441 acagctttta gaactggtta caatagcgac ggagagttag gttattggga taagttagag
     7501 ccactttata caatttttga tggtgtatct aaaacattct ctggtatttg gactcctgta
     7561 aagaatgact tcaaagagtt ttatgattta tacctttctg atgtagagaa atataatggt
     7621 tcggggaaat tgtttcccaa aacacctata cctgaaaatg ctttttctct ttctattatt
     7681 ccatggactt catttactgg gtttaactta aatatcaata ataatagtaa ttaccttcta
     7741 cccattatta cagcaggaaa attcattaat aaaggtaatt caatatattt accgctatct
     7801 ttacaggtac atcattctgt ttgtgatggt tatcatgcag gattgtttat gaactctatt
     7861 cagaattgtc agataggcct aatgactggc ttttataata tgagataatg ccgactgtac
     7921 tttttacagt cggttttcta atgtcactaa cctgccccgt tagttgatgc atgatcatca
     7981 taattctgtc tcattatcta gagtcgccat tggggaagaa gagcaggatt tgcctgcggt
     8041 tgaccgagac ggagtgggcg acgatgaacc ggatggcgtt cgcgtgcggt cagatggata
     8101 tctccatgga cacccacatg gatatctcca tggacaccca catggatatc tccatcaaat
     8161 cgaaggtaag caagctagac ggttcatgta catgaaccga acgcagcacc tccctcgcct
     8221 tgcggcgctg gagcaacaac ccgagtgccg aaggccgatg gaagaggaaa aagccgcaag
     8281 gcgagggagg tgctgcgttc gcgtttggga caacgtttag cgtcagtcga acagtggaat
     8341 ctcgccgggt agatgcttcg atttctccgc cgcctcgtcc tcgcgctcct tgcgagccct
     8401 gtagcagcgt tgacgcacgg caccctggct catgttgaaa tgctcggcgg tctgcggcca
     8461 tgtatggccc tcgtcatcgt gatatgagcg gactgcttcg cgcatagcgg cttgttcatc
     8521 aatccaatcc tcgcgcttca ttgccaccca gctctgaatg gtgcgcgtgg agacgccgta
     8581 tgcttcagct aattcacgtg cagacaccgg gcgcttcttc ctcaaagtcg tccttaccat
     8641 tccagtgcct ccttcatgac ctgctgatat ttgttctcac cgtgtttgtg tcctcgtgcg
     8701 gattggatgg cgatgaatgt ggctgcgttg gcaatggcac cgtcgagcca catgcggctg
     8761 cgggtgacga tccatttgtg gatgctcttg gcgatgtcct cgacctcacg cgcgtgcagc
     8821 ggatcgggga acagcgagac gttgagttcg tggcaggtgc ggcgcacgta taggcgcaat
     8881 gcctcgctgc tggcctccga acggtcgggc agccgccgca cgtagcggta ggcgagcgtg
     8941 cgggccttgt cgaagagcgt gcagttgcgt cccagcccct gcgtgcgggc gcgcttggtg
     9001 cgcttccagc tgacgggcgg catgtccccg tgttcctgga gcgactgcac gagctgttcc
     9061 aggtcgtagg tgtcccactc gatgatgtcc gacgcccacg ccggatgctc ggggttcttc
     9121 atcagaagtc ccgaatagcc catgtccccg tcgcaggagc ggcgcagtcc ctccgtggtg
     9181 gcgtgcagga gcttcaacgg cttgagacgc gccagatcgg ttctgggcac cggaaaggtg
     9241 agcacccagc ccgcgtgagc gtgcccgttg gccgggttct ccgcgatcca gttcggccgc
     9301 atgccctcgt gctcccacag ggccatcgcc ttggcgttcg cgtcgtcgat gtccaccacg
     9361 atcgtgttca ccagcgcggc ggggttcgcc tcgatgtgct tgcatttgag cgcgtcggcg
     9421 cggctcctgc ggtgcagctg ggagtagtcg tcgtcgcagc acagcgggcg ttcgggcagc
     9481 caccagcctt cgaacgtgcg cgacagctca agcgtcggct gcgaatactc atcggacatg
     9541 gctacagcat acatcgtatt tgtacagtga acaaatacgc gggaaccaag tcagatattt
     9601 cagcccagcg aagcgtcggc tgaacccagc ccagcacatc ctgcacgaaa agccgctgga
     9661 cggccctcag ccgcttctcc acgccgctca cgtatgcgcg agcctctctg acctgttcgg
     9721 ccgcgtggag gccactcacg gacgtttctg agccgtctga gccgtcggcg tccgctgcgc
     9781 gttccgcagg ctgaggc
//
