LOCUS       sequence-370917-       13674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NI-PE2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13674)
  AUTHORS   Velimirovic M, Zanetti LC, Shen MW, Fife JD, Lin L, Cha M, Akinci E,
            Barnum D, Yu T, Sherwood RI
  TITLE     Peptide fusion improves prime editing efficiency.
  JOURNAL   Nat Commun. 2022 Jun 18;13(1):3512. doi: 10.1038/s41467-022-31270-y.
  PUBMED    35717416
REFERENCE   2  (bases 1 to 13674)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2022-06-18"; volume: "13(1):3512. doi"; pages: " 
            10.1038/s41467-022-31270-y"
COMMENT     Sequence Label: p2T_CAG_SpCas9PE2_Igf1-NFATC2IP_BlastR (pepPE2_3)
FEATURES             Location/Qualifiers
     source          1..13674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     356..375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     589..611
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     603..620
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     604..620
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     630..649
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        630..648
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(679..690,691..853)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     regulatory      878..887
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(893..926)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        941..1320
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1600..2616
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2540..2562
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2624..2643
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3398..3418
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3443..5956,5957..5959,5960..7543)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3443..7543
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             9716..9736
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9745..9762
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(9785..9802)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9791..10015
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    10086..10220
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10110..10129)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     10164..10183
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(10378..10776)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(10795..10842)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(10890..11219)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10904..11039
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10958..10977)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     11204..11224
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     misc_feature    join(11234..11421,11422..11433)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(11490..11510)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11490..11508)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11529..11545)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11529..11545)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11542..11564)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11553..11569
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(11577..11583,11584..11601,11602..11607))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11622..11643
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11760..11777)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11931..12519)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12011..12030)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12690..13481,13482..13550))
                     /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     13313..13332
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13551..13655)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      241 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      301 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      361 agaaagcgaa aggagcgggc gctagggcgc gggcgggtgg ggtggtcacg ctgcgcgtaa
      421 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
      481 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
      541 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
      601 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
      661 ccggcatatg gttcttgaca gaggtgtaaa aagtactcaa aaattttact caagtgaaag
      721 tacaagtact tagggaaaat tttactcaat taaaagtaaa agtatctggc tagaatctta
      781 cttgagtaaa agtaaaaaag tactccatta aaattgtact tgagtattaa ggaagtaaaa
      841 gtaaaagcaa gaaagatcga tctcgaagga tctggaggcc accatggtgt cgataacttc
      901 gtatagcata cattatacga agttatcgat aagcttgctc gacattgatt attgactagt
      961 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     1021 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     1081 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     1141 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     1201 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     1261 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     1321 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
     1381 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg
     1441 gggcgcgcgc crgggggggg gggggggggs gggggggggg gsggggsgrg gsggrgrggk
     1501 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
     1561 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
     1621 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
     1681 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg
     1741 gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag
     1801 ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc
     1861 gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt
     1921 tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg
     1981 gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg
     2041 ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag
     2101 cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg
     2161 ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggrg
     2221 ggctcggggg rggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
     2281 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
     2341 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
     2401 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     2461 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
     2521 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
     2581 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
     2641 gtgctgtctc atcattttgg caaagaattc ctcgagcggc cgccagtgtg atggatatct
     2701 gcagaattcc agcacactgg cggccgttac tagtggatcc accggtacca tggctagcag
     2761 ggagggaatg aataagcctg aggggtacgg tgaggaggag cgccgcgcac ctcaggaggg
     2821 tattgtggac gagtgctgct tccgcgagtg cgacctgcgc cgcctcgaaa tgtactgtgc
     2881 ccctctcaag cctgccaaag aggctcgcga ggtgcgcgct cagcgccacg aggatatgcc
     2941 aaaagagcaa aaatatcaac ctcctgagga gaacaagaac gagaaagagc aacgccgcaa
     3001 aggggaggag tttggttcta caggaggtac tggaagtgcc gagccagtcg gtaaacgcgg
     3061 gcgctggtct ggcggctctg gcgcaggtcg cgggggtcgc ggtggctggg gcggtcgcgg
     3121 tcgccgccct cgcgctcaac gctccccaag ccgcggcacc ttggatgtgg tgtccgtgga
     3181 cctcgtgacc gactctgatg aggagatcct ggaggtggca actgcccgcg gcgccgcaga
     3241 tgaggttgag gtggagccac ctgagccacc agggcctgtt gctagccgcg actctagcgg
     3301 aggatctagc ggaggctcct ctggctctga gacacctggc acaagcgaga gcgcaacacc
     3361 tgaaagcagc gggggcagca gcggggggtc atgtacacca aagaagaagc ggaaggtcgg
     3421 tatccacgga gtcccagcag ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa
     3481 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
     3541 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
     3601 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
     3661 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
     3721 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
     3781 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
     3841 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
     3901 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
     3961 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     4021 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
     4081 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     4141 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     4201 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     4261 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     4321 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     4381 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     4441 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     4501 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     4561 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     4621 cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     4681 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     4741 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     4801 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     4861 cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg
     4921 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     4981 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     5041 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     5101 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     5161 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     5221 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     5281 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     5341 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     5401 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     5461 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     5521 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     5581 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     5641 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     5701 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     5761 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     5821 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     5881 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     5941 cgactacgat gtggacgcta tcgtgcctca gagctttctg aaggacgact ccatcgacaa
     6001 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     6061 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     6121 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     6181 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     6241 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     6301 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     6361 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     6421 aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa
     6481 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     6541 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     6601 cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
     6661 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
     6721 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag
     6781 gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
     6841 cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
     6901 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
     6961 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
     7021 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
     7081 ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
     7141 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
     7201 gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag
     7261 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta
     7321 caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac
     7381 cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa
     7441 gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
     7501 cctgtacgag acacggatcg acctgtctca gctgggaggt gactctggag gatctagcgg
     7561 aggatcctct ggcagcgaga caccaggaac aagcgagtca gcaacaccag agagcagtgg
     7621 cggcagcagc ggcggcagca gcaccctaaa tatagaagat gagtatcggc tacatgagac
     7681 ctcaaaagag ccagatgttt ctctagggtc cacatggctg tctgattttc ctcaggcctg
     7741 ggcggaaacc gggggcatgg gactggcagt tcgccaagct cctctgatca tacctctgaa
     7801 agcaacctct acccccgtgt ccataaaaca ataccccatg tcacaagaag ccagactggg
     7861 gatcaagccc cacatacaga gactgttgga ccagggaata ctggtaccct gccagtcccc
     7921 ctggaacacg cccctgctac ccgttaagaa accagggact aatgattata ggcctgtcca
     7981 ggatctgaga gaagtcaaca agcgggtgga agacatccac cccaccgtgc ccaaccctta
     8041 caacctcttg agcgggctcc caccgtccca ccagtggtac actgtgcttg atttaaagga
     8101 tgcctttttc tgcctgagac tccaccccac cagtcagcct ctcttcgcct ttgagtggag
     8161 agatccagag atgggaatct caggacaatt gacctggacc agactcccac agggtttcaa
     8221 aaacagtccc accctgttta atgaggcact gcacagagac ctagcagact tccggatcca
     8281 gcacccagac ttgatcctgc tacagtacgt ggatgactta ctgctggccg ccacttctga
     8341 gctagactgc caacaaggta ctcgggccct gttacaaacc ctagggaacc tcgggtatcg
     8401 ggcctcggcc aagaaagccc aaatttgcca gaaacaggtc aagtatctgg ggtatcttct
     8461 aaaagagggt cagagatggc tgactgaggc cagaaaagag actgtgatgg ggcagcctac
     8521 tccgaagacc cctcgacaac taagggagtt cctagggaag gcaggcttct gtcgcctctt
     8581 catccctggg tttgcagaaa tggcagcccc cctgtaccct ctcaccaaac cggggactct
     8641 gtttaattgg ggcccagacc aacaaaaggc ctatcaagaa atcaagcaag ctcttctaac
     8701 tgccccagcc ctggggttgc cagatttgac taagcccttt gaactctttg tcgacgagaa
     8761 gcagggctac gccaaaggtg tcctaacgca aaaactggga ccttggcgtc ggccggtggc
     8821 ctacctgtcc aaaaagctag acccagtagc agctgggtgg cccccttgcc tacggatggt
     8881 agcagccatt gccgtactga caaaggatgc aggcaagcta accatgggac agccactagt
     8941 cattctggcc ccccatgcag tagaggcact agtcaaacaa ccccccgacc gctggctttc
     9001 caacgcccgg atgactcact atcaggcctt gcttttggac acggaccggg tccagttcgg
     9061 accggtggta gccctgaacc cggctacgct gctcccactg cctgaggaag ggctgcaaca
     9121 caactgcctt gatatcctgg ccgaagccca cggaacccga cccgacctaa cggaccagcc
     9181 gctcccagac gccgaccaca cctggtacac ggatggaagc agtctcttac aagagggaca
     9241 gcgtaaggcg ggagctgcgg tgaccaccga gaccgaggta atctgggcta aagccctgcc
     9301 agccgggaca tccgctcagc gggctgaact gatagcactc acccaggccc taaagatggc
     9361 agaaggtaag aagctaaatg tttatactga tagccgttat gcttttgcta ctgcccatat
     9421 ccatggagaa atatacagaa ggcgtgggtg gctcacatca gaaggcaaag agatcaaaaa
     9481 taaagacgag atcttggccc tactaaaagc cctctttctg cccaaaagac ttagcataat
     9541 ccattgtcca ggacatcaaa agggacacag cgccgaggct agaggcaacc ggatggctga
     9601 ccaagcggcc cgaaaggcag ccatcacaga gactccagac acctctaccc tcctcataga
     9661 aaattcatca ccctctggcg gctcaaaaag aaccgccgac ggcagcgaat tcgagcccaa
     9721 gaagaagagg aaagtctaac cggtcatcat caccatcacc attgagttta aacccgctga
     9781 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     9841 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     9901 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     9961 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
    10021 gaggcggaaa gaacaagatc tagcggccgc caccgcggtg gagctcgaat taattcatcg
    10081 atgatgatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
    10141 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
    10201 taagctgcaa taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgg
    10261 aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg
    10321 gcggtggaat cgaaatctcg tagcacgtgt cagtcctgct cctcggccac gaagtgctta
    10381 gccctcccac acataaccag agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc
    10441 atcactgtcc ttcactatgg ctttgatccc aggatgcaga tcgagaagca cctgtcggca
    10501 ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg atcgcgacga tacaagtcag
    10561 gttgccagct gccgcagcag cagcagtgcc cagcaccacg agttctgcac aaggtccccc
    10621 agtaaaatga tatacattga caccagtgaa gatgcggccg tcgctagaga gagctgcgct
    10681 ggcgacgctg tagtcttcag agatggggat gctgttgatt gtagccgttg ctctttcaat
    10741 gagggtggat tcttcttgag acaaaggctt ggccatggtt tagttcctca ccttgtcgta
    10801 ttatactatg ccgatatact atgccgatga ttaattgtca acacgtgctg atcagatccg
    10861 aaaatggata tacaagctcc cgggagcttt ttgcaaaagc ctaggcctcc aaaaaagcct
    10921 cctcactact tctggaatag ctcagaggca gaggcggcct cggcctctgc ataaataaaa
    10981 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
    11041 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
    11101 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
    11161 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
    11221 ttccacagaa ttcaagtgat ctccaaaaaa taagtacttt ttgactgtaa ataaaattgt
    11281 aaggagtaaa aagtactttt ttttctaaaa aaatgtaatt aagtaaaagt aaaagtattg
    11341 atttttaatt gtactcaagt aaagtaaaaa tccccaaaaa taatacttaa gtacagtaat
    11401 caagtaaaat tactcaagta ctttacacct ctggttcttg accccctacc ttcagcaagc
    11461 ccagcagatc cgagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
    11521 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
    11581 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
    11641 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
    11701 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
    11761 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
    11821 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
    11881 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
    11941 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
    12001 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
    12061 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
    12121 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
    12181 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
    12241 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
    12301 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
    12361 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
    12421 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
    12481 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
    12541 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
    12601 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
    12661 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
    12721 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
    12781 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
    12841 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
    12901 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
    12961 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
    13021 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
    13081 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
    13141 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
    13201 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
    13261 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
    13321 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
    13381 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
    13441 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
    13501 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
    13561 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
    13621 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtg
//
