LOCUS       sequence-388916-       14235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpRY-YE1-BE4max and blasticidin 
            resistance: EFS promoter-SpRY-YE1-BE4max-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14235)
  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 14235)
  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: SpRY-YE1-BE4max
FEATURES             Location/Qualifiers
     source          1..14235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        181..560
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        562..760
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     715..735
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             778..958
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1005..1130
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1623..1856
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2041..2085
                     /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    2383..2500
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2593..2804
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2855..2875
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3656..7756
                     /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
                     LIGALLFDSGETAERTRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SIRPKRNSDKLIARKKDWDPKKYGGFLWPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAKQLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTRLGAPRAFKYFDTTIDPKQYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7787..8035
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8066..8314
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8357..8377
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8378..8401
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8411..8467
                     /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             8468..8866
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8891..9479
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8944..8964)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9362..9373)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9482..9498)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9483..9499)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10004..10184
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10210..10227)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10216..10440
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10486..10914
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10573..10592)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10783..10804
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10923..10943)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      11088..11222
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11149..11168
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11284..11331
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11350..11724
                     /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    11854..11975
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11891..11910)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11945..11964
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12024..12040)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12024..12040)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12037..12059)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12048..12064
                     /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(12072..12078,12079..12096,12097..12102))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12117..12138
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12255..12272)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12426..13014)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12506..12525)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13185..13976,13977..14045))
                     /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     13808..13827
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14046..14150)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(14225..14235,1..9))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
       61 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
      121 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
      181 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      241 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      301 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      361 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      421 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      481 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      541 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      601 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      661 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      721 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      781 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      841 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      901 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      961 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
     1021 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
     1081 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
     1141 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1201 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1261 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1321 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1381 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1441 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1501 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1561 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1621 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1681 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1741 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1801 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1861 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1921 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1981 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     2041 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     2101 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     2161 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2221 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2281 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2341 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2401 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2461 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2521 cagagatcca gtttggttaa ttagctagct aggtcttgaa aggagtggga attggctccg
     2581 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
     2641 tcggcaattg atccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
     2701 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
     2761 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggaccgg ttgccaccat
     2821 gaaacggaca gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtctcctc
     2881 agagactggg cctgtcgccg tcgatccaac cctgcgccgc cggattgaac ctcacgagtt
     2941 tgaagtgttc tttgaccccc gggagctgag aaaggagaca tgcctgctgt acgagatcaa
     3001 ctggggaggc aggcactcca tctggaggca cacctctcag aacacaaata agcacgtgga
     3061 ggtgaacttc atcgagaagt ttaccacaga gcggtacttc tgccccaata ccagatgtag
     3121 catcacatgg tttctgagct attccccttg cggagagtgt agcagggcca tcaccgagtt
     3181 cctgtccaga tatccacacg tgacactgtt tatctacatc gccaggctgt atcaccacgc
     3241 agacccagag aataggcagg gcctgcgcga tctgatcagc tccggcgtga ccatccagat
     3301 catgacagag caggagtccg gctactgctg gcggaacttc gtgaattatt ctcctagcaa
     3361 cgaggcccac tggcctaggt acccacacct gtgggtgcgc ctgtacgtgc tggagctgta
     3421 ttgcatcatc ctgggcctgc ccccttgtct gaatatcctg cggagaaagc agccccagct
     3481 gaccttcttt acaatcgccc tgcagtcttg tcactatcag aggctgccac cccacatcct
     3541 gtgggccaca ggcctgaagt ctggaggatc tagcggagga tcctctggca gcgagacacc
     3601 aggaacaagc gagtcagcaa caccagagag cagtggcggc agcagcggcg gcagcgacaa
     3661 gaagtacagc atcggcctgg ccatcggcac caactctgtg ggctgggccg tgatcaccga
     3721 cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat
     3781 caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg agcggacccg
     3841 gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca
     3901 agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga
     3961 gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt
     4021 ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt
     4081 ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa
     4141 gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa
     4201 gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc
     4261 cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga
     4321 aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc
     4381 cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa
     4441 actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg
     4501 cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag
     4561 cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa
     4621 gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct
     4681 gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat
     4741 tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat
     4801 ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg
     4861 gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct
     4921 gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat
     4981 cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc
     5041 ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga
     5101 caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc
     5161 caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga
     5221 gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga
     5281 gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca
     5341 gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt
     5401 ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga
     5461 caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct
     5521 gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt
     5581 cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag
     5641 ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct
     5701 gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac
     5761 ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca
     5821 cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt
     5881 ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat
     5941 ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg
     6001 gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa
     6061 cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta
     6121 cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc
     6181 tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa
     6241 ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg
     6301 gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc
     6361 cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga
     6421 aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta
     6481 cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt
     6541 gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca
     6601 cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa
     6661 gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc
     6721 caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat
     6781 gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat
     6841 cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt
     6901 gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg
     6961 cggcttcagc aaagagtcta tccggcccaa gaggaacagc gataagctga tcgccagaaa
     7021 gaaggactgg gaccctaaga agtacggcgg cttcctgtgg cccaccgtgg cctattctgt
     7081 gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct
     7141 actggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga
     7201 agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct
     7261 gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccaagcagc tgcagaaggg
     7321 aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga
     7381 gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa
     7441 gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc
     7501 cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag
     7561 agagcaggcc gagaatatca tccacctgtt taccctgacc cggctgggag cccctcgggc
     7621 cttcaagtac tttgacacca ccatcgaccc caagcagtac cggagcacca aagaggtgct
     7681 ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc
     7741 tcagctggga ggcgacagcg gcgggagcgg cgggagcggg gggagcacta atctgagcga
     7801 catcattgag aaggagactg ggaaacagct ggtcattcag gagtccatcc tgatgctgcc
     7861 tgaggaggtg gaggaagtga tcggcaacaa gccagagtct gacatcctgg tgcacaccgc
     7921 ctacgacgag tccacagatg agaatgtgat gctgctgacc tctgacgccc ccgagtataa
     7981 gccttgggcc ctggtcatcc aggattctaa cggcgagaat aagatcaaga tgctgagcgg
     8041 aggatccgga ggatctggag gcagcaccaa cctgtctgac atcatcgaga aggagacagg
     8101 caagcagctg gtcatccagg agagcatcct gatgctgccc gaagaagtcg aagaagtgat
     8161 cggaaacaag cctgagagcg atatcctggt ccataccgcc tacgacgaga gtaccgacga
     8221 aaatgtgatg ctgctgacat ccgacgcccc agagtataag ccctgggctc tggtcatcca
     8281 ggattccaac ggagagaaca aaatcaaaat gctgtctggc ggctcaaaaa gaaccgccga
     8341 cggcagcgaa ttcgagccca agaagaagag gaaagtcgat tacaaagacg atgacgataa
     8401 gggatccggc gcaacaaact tctctctgct gaaacaagcc ggagatgtcg aagagaatcc
     8461 tggaccgatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc
     8521 tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag
     8581 cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga
     8641 actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc
     8701 gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct
     8761 cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt
     8821 tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaagaat tcgatatcaa
     8881 gcttatcggt aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     8941 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     9001 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     9061 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     9121 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     9181 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     9241 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     9301 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     9361 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     9421 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     9481 tcgataccgt cgacctcgag acctagaaaa acatggagca atcacaagta gcaatacagc
     9541 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc
     9601 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     9661 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatat
     9721 ccttgatctg tggatctacc acacacaagg ctacttccct gattggcaga actacacacc
     9781 agggccaggg atcagatatc cactgacctt tggatggtgc tacaagctag taccagttga
     9841 gcaagagaag gtagaagaag ccaatgaagg agagaacacc cgcttgttac accctgtgag
     9901 cctgcatggg atggatgacc cggagagaga agtattagag tggaggtttg acagccgcct
     9961 agcatttcat cacatggccc gagagctgca tccggactgt actgggtctc tctggttaga
    10021 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
    10081 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
    10141 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttaaaccc
    10201 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
    10261 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
    10321 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
    10381 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
    10441 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
    10501 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
    10561 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
    10621 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
    10681 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
    10741 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
    10801 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
    10861 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
    10921 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
    10981 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
    11041 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
    11101 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
    11161 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
    11221 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca
    11281 cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
    11341 aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc
    11401 ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac
    11461 ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg
    11521 gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag
    11581 tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc
    11641 ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac
    11701 ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc
    11761 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
    11821 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
    11881 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
    11941 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
    12001 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
    12061 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
    12121 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
    12181 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
    12241 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
    12301 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
    12361 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
    12421 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
    12481 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
    12541 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
    12601 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
    12661 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
    12721 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
    12781 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
    12841 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
    12901 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
    12961 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
    13021 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
    13081 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
    13141 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
    13201 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
    13261 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
    13321 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
    13381 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
    13441 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
    13501 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
    13561 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
    13621 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
    13681 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
    13741 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
    13801 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
    13861 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
    13921 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
    13981 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
    14041 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
    14101 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
    14161 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
    14221 tcagtacaat ctgct
//
