LOCUS       sequence-388909-       14391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-NG-CGBE1 and blasticidin 
            resistance: EFS promoter-SpCas9-NG-CGBE1-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14391)
  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 14391)
  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-NG-CGBE1
FEATURES             Location/Qualifiers
     source          1..14391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        187..566
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        568..766
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     721..741
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             784..964
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1011..1136
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1629..1862
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2047..2091
                     /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    2389..2506
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2599..2810
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2861..2881
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4376..8476
                     /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
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SIRPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARFLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPRAFKYFDTTIDRKVYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8519..8539
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8540..8563
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8573..8629
                     /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             8630..9028
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9053..9641
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9106..9126)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9524..9535)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9644..9660)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9645..9661)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10166..10346
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10372..10389)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10378..10602
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10648..11076
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10735..10754)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10945..10966
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11085..11105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      11250..11384
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11311..11330
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11446..11493
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11512..11886
                     /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    12016..12137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12053..12072)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12107..12126
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12186..12202)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12186..12202)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12199..12221)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12210..12226
                     /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(12234..12240,12241..12258,12259..12264))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12279..12300
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12417..12434)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12588..13176)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12668..12687)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13347..14138,14139..14207))
                     /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     13970..13989
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14208..14312)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(14387..14391,1..15))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
       61 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      121 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      181 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      241 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      301 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      361 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      421 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      481 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      541 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      601 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      661 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      721 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
      781 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
      841 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
      901 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
      961 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     1021 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     1081 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     1141 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     1201 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     1261 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     1321 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     1381 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     1441 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
     1501 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     1561 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     1621 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     1681 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     1741 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     1801 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     1861 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     1921 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     1981 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     2041 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     2101 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     2161 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     2221 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     2281 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg
     2341 gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa
     2401 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     2461 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     2521 ggacagcaga gatccagttt ggttaattag ctagctaggt cttgaaagga gtgggaattg
     2581 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     2641 gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     2701 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     2761 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gaccggttgc
     2821 caccatgaaa cggacagccg acggaagcga gttcgagtca ccaaagaaga agcggaaagt
     2881 cgccaacgaa ctgacctggc acgacgtgct ggccgaagag aagcagcagc cctatttcct
     2941 taacaccctg cagaccgtgg ccagcgagcg gcagagcggc gtgaccatct accccccaca
     3001 gaaagacgtg tttaacgcct tccgcttcac agagctgggc gacgtgaagg tggtgatcct
     3061 gggccaggac ccctaccacg gccccggcca ggcccatggt ctggccttca gcgtgcggcc
     3121 cggcatcgcc atccccccca gcctgctgaa catgtataag gagctggaaa acaccatccc
     3181 cggcttcacc cggcccaatc acggctacct ggagagctgg gcgcggcagg gcgtgctgct
     3241 gctcaacacc gtgctgacgg tacgggccgg ccaggcgcat agccacgcca gcctgggctg
     3301 ggagaccttc accgataagg tgatcagcct gatcaaccag caccgggagg gcgtggtgtt
     3361 tctgctgtgg ggcagccatg cccagaagaa aggcgccatc atcgacaagc agcggcatca
     3421 tgtgctgaaa gccccccacc ccagccccct tagcgcccac cggggcttct tcggctgcaa
     3481 ccatttcgtg ctggccaatc agtggctgga acaacggggc gagaccccca ttgactggat
     3541 gcccgtgttg cccgccgaga gcgagagcgg cgggagcggc gggagcgggg ggagctcctc
     3601 agagactggg cctgtcgccg tcgatccaac cctgcgccgc cggattgaac ctcacgagtt
     3661 tgaagtgttc tttgaccccc gggagctggc caaggagaca tgcctgctgt acgagatcaa
     3721 ctggggaggc aggcactcca tctggaggca cacctctcag aacacaaata agcacgtgga
     3781 ggtgaacttc atcgagaagt ttaccacaga gcggtacttc tgccccaata ccagatgtag
     3841 catcacatgg tttctgagct ggtccccttg cggagagtgt agcagggcca tcaccgagtt
     3901 cctgtccaga tatccacacg tgacactgtt tatctacatc gccaggctgt atcaccacgc
     3961 agacccaagg aataggcagg gcctgcgcga tctgatcagc tccggcgtga ccatccagat
     4021 catgacagag caggagtccg gctactgctg gcggaacttc gtgaattatt ctcctagcaa
     4081 cgaggcccac tggcctaggt acccacacct gtgggtgcgc ctgtacgtgc tggagctgta
     4141 ttgcatcatc ctgggcctgc ccccttgtct gaatatcctg cggagaaagc agccccagct
     4201 gaccttcttt acaatcgccc tgcagtcttg tcactatcag aggctgccac cccacatcct
     4261 gtgggccaca ggcctgaagt ctggaggatc tagcggagga tcctctggca gcgagacacc
     4321 aggaacaagc gagtcagcaa caccagagag cagtggcggc agcagcggcg gcagcgacaa
     4381 gaagtacagc atcggcctgg ccatcggcac caactctgtg ggctgggccg tgatcaccga
     4441 cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat
     4501 caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg
     4561 gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca
     4621 agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga
     4681 gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt
     4741 ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt
     4801 ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa
     4861 gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa
     4921 gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc
     4981 cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga
     5041 aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc
     5101 cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa
     5161 actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg
     5221 cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag
     5281 cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa
     5341 gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct
     5401 gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat
     5461 tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat
     5521 ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg
     5581 gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct
     5641 gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat
     5701 cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc
     5761 ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga
     5821 caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc
     5881 caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga
     5941 gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga
     6001 gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca
     6061 gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt
     6121 ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga
     6181 caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct
     6241 gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt
     6301 cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag
     6361 ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct
     6421 gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac
     6481 ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca
     6541 cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt
     6601 ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat
     6661 ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg
     6721 gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa
     6781 cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta
     6841 cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc
     6901 tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa
     6961 ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg
     7021 gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc
     7081 cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga
     7141 aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta
     7201 cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt
     7261 gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca
     7321 cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa
     7381 gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc
     7441 caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat
     7501 gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat
     7561 cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt
     7621 gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg
     7681 cggcttcagc aaagagtcta tccggcccaa gaggaacagc gataagctga tcgccagaaa
     7741 gaaggactgg gaccctaaga agtacggcgg cttcgtgagc cccaccgtgg cctattctgt
     7801 gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct
     7861 actggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga
     7921 agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct
     7981 gttcgagctg gaaaacggcc ggaagagaat gctggcctct gcccggttcc tgcagaaggg
     8041 aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga
     8101 gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa
     8161 gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc
     8221 cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag
     8281 agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag cccctcgggc
     8341 cttcaagtac tttgacacca ccatcgaccg gaaggtgtac cggagcacca aagaggtgct
     8401 ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc
     8461 tcagctggga ggcgactctg gcggctcaaa aagaaccgcc gacggcagcg aattcgagcc
     8521 caagaagaag aggaaagtcg attacaaaga cgatgacgat aagggatccg gcgcaacaaa
     8581 cttctctctg ctgaaacaag ccggagatgt cgaagagaat cctggaccga tggccaagcc
     8641 tttgtctcaa gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc
     8701 catctctgaa gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac
     8761 tggtgtcaat gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac
     8821 tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag
     8881 gggcatcttg agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat
     8941 caaagccata gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct
     9001 gccctctggt tatgtgtggg agggctaaga attcgatatc aagcttatcg gtaatcaacc
     9061 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     9121 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     9181 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     9241 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     9301 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     9361 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     9421 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     9481 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     9541 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     9601 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg
     9661 agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg
     9721 tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc
     9781 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     9841 gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc tgtggatcta
     9901 ccacacacaa ggctacttcc ctgattggca gaactacaca ccagggccag ggatcagata
     9961 tccactgacc tttggatggt gctacaagct agtaccagtt gagcaagaga aggtagaaga
    10021 agccaatgaa ggagagaaca cccgcttgtt acaccctgtg agcctgcatg ggatggatga
    10081 cccggagaga gaagtattag agtggaggtt tgacagccgc ctagcatttc atcacatggc
    10141 ccgagagctg catccggact gtactgggtc tctctggtta gaccagatct gagcctggga
    10201 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
    10261 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
    10321 ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac ccgctgatca gcctcgactg
    10381 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
    10441 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
    10501 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
    10561 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
    10621 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
    10681 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
    10741 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
    10801 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
    10861 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
    10921 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
    10981 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
    11041 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
    11101 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
    11161 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
    11221 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca gttccgccca
    11281 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
    11341 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
    11401 gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca
    11461 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag
    11521 ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg
    11581 accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg
    11641 gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg
    11701 gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc
    11761 acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg
    11821 cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag
    11881 gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
    11941 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
    12001 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
    12061 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
    12121 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
    12181 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
    12241 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
    12301 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
    12361 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
    12421 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
    12481 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
    12541 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
    12601 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
    12661 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
    12721 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
    12781 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
    12841 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
    12901 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
    12961 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
    13021 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
    13081 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
    13141 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
    13201 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
    13261 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
    13321 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
    13381 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
    13441 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
    13501 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
    13561 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
    13621 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
    13681 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
    13741 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
    13801 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
    13861 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
    13921 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
    13981 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
    14041 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
    14101 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
    14161 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
    14221 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
    14281 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
    14341 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca a
//
