LOCUS       sequence-388895-       14301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-NRCH-APOBEC-nCas9-Ung and 
            blasticidin resistance: EFS 
            promoter-SpCas9-NRCH-APOBEC-nCas9-Ung-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14301)
  AUTHORS   Kim N, Choi S, Kim S, Song M, Seo JH, Min S, Park J, Cho SR, Kim HH
  TITLE     Deep learning models to predict the editing efficiencies and 
            outcomes of diverse base editors.
  JOURNAL   Nat Biotechnol. 2024 Mar;42(3):484-497. doi: 
            10.1038/s41587-023-01792-x
  PUBMED    37188916
REFERENCE   2  (bases 1 to 14301)
  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"; doi: "10"; journalName: 
            "Nat Biotechnol"; date: "2024-03"; volume: "42"; issue: "3"; pages: 
            "484-497"
COMMENT     Sequence Label: SpCas9-NRCH-APOBEC-nCas9-Ung
FEATURES             Location/Qualifiers
     source          1..14301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(34..53)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        225..604
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        606..804
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     759..779
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             822..1002
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1049..1174
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1667..1900
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2085..2129
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2427..2544
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2637..2848
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2866..2886
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2887..3570
                     /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="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
                     RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
                     RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
                     AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
                     WATGLK"
     CDS             3622..7722
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMVKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQGDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRLRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGGH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKGNSDKLIARKKDWDPKKYGGFNSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTINRKQYNTTKEVLD
                     ATLIRQSITGLYETRIDLSQLGGD"
     CDS             7723..7770
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8467..8487
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8488..8511
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8521..8577
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             8578..8976
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9001..9589
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9054..9074)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9472..9483)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9592..9608)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9593..9609)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10114..10294
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10320..10337)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10326..10550
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10596..11024
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10683..10702)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10893..10914
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11033..11053)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      11198..11332
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11259..11278
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11394..11441
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11460..11834
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    11964..12085
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12001..12020)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12055..12074
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12134..12150)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12134..12150)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12147..12169)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12158..12174
                     /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(12182..12188,12189..12206,12207..12212))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12227..12248
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12365..12382)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12536..13124)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12616..12635)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13295..14086,14087..14155))
                     /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     13918..13937
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14156..14260)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
       61 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
      121 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
      181 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
      241 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      301 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      361 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      421 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      481 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      541 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      601 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      661 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      721 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      781 acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc
      841 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
      901 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
      961 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     1021 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     1081 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     1141 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     1201 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     1261 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     1321 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     1381 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     1441 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     1501 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     1561 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1621 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     1681 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     1741 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     1801 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     1861 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     1921 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     1981 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     2041 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     2101 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     2161 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     2221 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     2281 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     2341 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     2401 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     2461 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     2521 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     2581 ttaattagct agctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc
     2641 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgatccgg
     2701 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     2761 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     2821 tcgcaacggg tttgccgcca gaacacagga ccggttgcca ccatgccaaa gaagaagagg
     2881 aaggtttcat cggagaccgg ccctgttgct gttgacccca ccctgcggcg gagaatcgag
     2941 ccacacgagt tcgaggtgtt cttcgaccca agggagctcc gcaaggagac gtgcctcctg
     3001 tacgagatca actggggcgg caggcactcc atctggaggc acaccagcca aaacaccaac
     3061 aagcacgtgg aggtcaactt catcgagaag ttcaccaccg agaggtactt ctgcccaaac
     3121 acccgctgct ccatcacctg gttcctgtcc tggagcccat gcggcgagtg ctccagggcc
     3181 atcaccgagt tcctcagccg ctacccacac gtcaccctgt tcatctacat cgccaggctc
     3241 taccaccacg ccgacccaag gaacaggcag ggcctccgcg acctgatctc cagcggcgtg
     3301 accatccaaa tcatgaccga gcaggagtcc ggctactgct ggaggaactt cgtcaactac
     3361 tccccaagca acgaggccca ctggccaagg tacccacacc tctgggtgcg cctctacgtg
     3421 ctcgagctgt actgcatcat cctcggcctg ccaccatgcc tcaacatcct gaggcgcaag
     3481 caaccacagc tgaccttctt caccatcgcc ctccaaagct gccactacca gaggctccca
     3541 ccacacatcc tgtgggctac cggcctcaag tccggcagcg agacgccagg cacctccgag
     3601 agcgctacgc ctgaacttaa ggacaagaag tacagcatcg gcctggccat cggcaccaac
     3661 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
     3721 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac
     3781 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
     3841 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
     3901 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
     3961 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
     4021 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
     4081 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
     4141 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     4201 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     4261 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
     4321 aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
     4381 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
     4441 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
     4501 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
     4561 gcccccctga gcgcctctat ggtgaagaga tacgacgagc accaccagga cctgaccctg
     4621 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
     4681 agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag
     4741 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
     4801 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcatcatccc ccaccagatc
     4861 cacctgggag agctgcacgc cattctgcgg cggcagggcg atttttaccc attcctgaag
     4921 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
     4981 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
     5041 ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg
     5101 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
     5161 tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg
     5221 agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag
     5281 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
     5341 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
     5401 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
     5461 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
     5521 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggctg
     5581 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
     5641 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
     5701 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
     5761 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
     5821 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggcggccac
     5881 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
     5941 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
     6001 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
     6061 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
     6121 gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac
     6181 aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag
     6241 gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag
     6301 agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
     6361 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
     6421 ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa
     6481 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
     6541 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
     6601 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
     6661 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
     6721 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
     6781 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg
     6841 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc
     6901 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagggc
     6961 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
     7021 aacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
     7081 aagaaactga agagtgtgaa agagctactg gggatcacca tcatggaaag aagcagcttc
     7141 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
     7201 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
     7261 gcctctgccg gcgtgctgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
     7321 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
     7381 aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc
     7441 gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac
     7501 aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc
     7561 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat caaccggaag
     7621 cagtacaaca ccaccaaaga ggtgctggac gccaccctga tccggcagag catcaccggc
     7681 ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acaagagacc cgcagcaacc
     7741 aagaaggcag ggcaagcaaa gaagaagaag acgcgtgact ccggcggcag catgtttgga
     7801 gagagctgga agaagcacct cagcggggag ttcgggaaac cgtattttat caagctaatg
     7861 ggatttgttg cagaagaaag aaagcattac actgtttatc cacccccaca ccaagtcttc
     7921 acctggaccc agatgtgtga cataaaagat gtgaaggttg tcatcctggg acaggatcca
     7981 tatcatggac ctaatcaagc tcacgggctc tgctttagtg ttcaaaggcc tgttccgcct
     8041 ccgcccagtt tggagaacat ttataaagag ttgtctacag acatagagga ttttgttcat
     8101 cctggccatg gagatttatc tgggtgggcc aagcaaggtg ttctccttct caacgctgtc
     8161 ctcacggttc gtgcccatca agccaactct cataaggagc gaggctggga gcagttcact
     8221 gatgcagttg tgtcctggct aaatcagaac tcgaatggcc ttgttttctt gctctggggc
     8281 tcttatgctc agaagaaggg cagtgccatt gataggaagc ggcaccatgt actacagacg
     8341 gctcatccct cccctttgtc agtgtataga gggttctttg gatgtagaca cttttcaaag
     8401 accaatgagc tgctgcagaa gtctggcaag aagcccattg actggaagga gctgtcgggg
     8461 gggagcccaa agaagaagcg gaaggtggat tacaaagacg atgacgataa gggatccggc
     8521 gcaacaaact tctctctgct gaaacaagcc ggagatgtcg aagagaatcc tggaccgatg
     8581 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
     8641 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
     8701 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
     8761 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
     8821 gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat
     8881 cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
     8941 gaattgctgc cctctggtta tgtgtgggag ggctaagaat tcgatatcaa gcttatcggt
     9001 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     9061 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     9121 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     9181 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     9241 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     9301 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     9361 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     9421 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     9481 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     9541 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt
     9601 cgacctcgag acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat
     9661 gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct
     9721 caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     9781 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg
     9841 tggatctacc acacacaagg ctacttccct gattggcaga actacacacc agggccaggg
     9901 atcagatatc cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag
     9961 gtagaagaag ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg
    10021 atggatgacc cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat
    10081 cacatggccc gagagctgca tccggactgt actgggtctc tctggttaga ccagatctga
    10141 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
    10201 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
    10261 agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttaaaccc gctgatcagc
    10321 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
    10381 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
    10441 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
    10501 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
    10561 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
    10621 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
    10681 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
    10741 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
    10801 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
    10861 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
    10921 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
    10981 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
    11041 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
    11101 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
    11161 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccagt
    11221 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
    11281 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
    11341 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca
    11401 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca
    11461 tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg
    11521 agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg
    11581 tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca
    11641 acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg
    11701 tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc
    11761 cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg
    11821 aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt
    11881 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
    11941 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
    12001 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
    12061 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
    12121 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
    12181 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
    12241 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
    12301 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
    12361 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
    12421 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
    12481 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
    12541 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
    12601 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
    12661 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
    12721 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
    12781 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
    12841 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
    12901 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
    12961 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
    13021 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
    13081 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
    13141 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
    13201 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
    13261 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
    13321 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
    13381 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
    13441 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
    13501 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
    13561 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
    13621 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
    13681 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
    13741 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
    13801 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
    13861 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
    13921 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
    13981 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
    14041 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
    14101 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
    14161 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
    14221 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
    14281 cacctgacgt cgacggatcg g
//
