LOCUS       sequence-372774-        9270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding pCas9 and gRNA for mutagenesis or gene-correction 
            of human ABCA3 mutation at amino acid W308.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9270)
  AUTHORS   Sun YL, Hennessey EE, Heins H, Yang P, Villacorta-Martin C, Kwan J, 
            Gopalan K, James M, Emili A, Cole FS, Wambach JA, Kotton DN
  TITLE     Human pluripotent stem cell modeling of alveolar type 2 cell 
            dysfunction caused by ABCA3 mutations.
  JOURNAL   J Clin Invest. 2024 Jan 16;134(2):e164274. doi: 10.1172/JCI164274.
  PUBMED    38226623
REFERENCE   2  (bases 1 to 9270)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Clin 
            Invest."; date: "2024-01-16"; volume: "134(2):e164274. doi"; pages: 
            " 10.1172/JCI164274"
COMMENT     Sequence Label: pX330-U6-W308_guide-CBh-hSpCas9
FEATURES             Location/Qualifiers
     source          1..9270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        271..346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        443..728
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        730..986
                     /label=chicken beta-actin promoter
                     /note="contains a 21-bp deletion relative to the standard 
                     chicken beta-actin promoter"
     intron          987..1214
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1226..1235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1235..1300
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1307..1327
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1352..5452
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1352..5452
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5453..5500
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5516..5569
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(5570..5572,5573..6283)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5570..5572,5573..6283)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5873..5892)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6220..6241
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6311..6328)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6317..6524
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6533..6673
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6533..6662
                     /label=AAV2 ITR
     rep_origin      6748..7203
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6835..6854)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7045..7066
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7220..7239)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7339..7361
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7399..7417)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7485..7589
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7590..7658,7659..8450)
                     /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     complement(7808..7827)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8621..9209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9110..9129
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg gaggaagaac aagaggaacc gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
      361 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
      421 gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg cctggctgac
      481 cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact ttccattgac
      541 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      601 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattgtgccc
      661 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      721 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
      781 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
      841 gggggggggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca
      901 atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct
      961 ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc
     1021 gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg
     1081 tgagcgggcg ggacggccct tctcctccgg gctgtaatta gctgagcaag aggtaagggt
     1141 ttaagggatg gttggttggt ggggtattaa tgtttaatta cctggagcac ctgcctgaaa
     1201 tcactttttt tcaggttgga ccggtgccac catggactat aaggaccacg acggagacta
     1261 caaggatcat gatattgatt acaaagacga tgacgataag atggccccaa agaagaagcg
     1321 gaaggtcggt atccacggag tcccagcagc cgacaagaag tacagcatcg gcctggacat
     1381 cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa
     1441 attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct
     1501 gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag
     1561 atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc
     1621 caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa
     1681 gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa
     1741 gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct
     1801 gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga
     1861 gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac
     1921 ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat
     1981 cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg
     2041 cgagaagaag aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa
     2101 cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta
     2161 cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct
     2221 ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga
     2281 gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga
     2341 cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt
     2401 cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga
     2461 gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt
     2521 gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc
     2581 ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc
     2641 attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta
     2701 cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga
     2761 aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt
     2821 catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca
     2881 cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac
     2941 cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct
     3001 gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa
     3061 aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct
     3121 gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga
     3181 aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat
     3241 gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct
     3301 gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg
     3361 ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag
     3421 aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc
     3481 ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc
     3541 cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat
     3601 gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca
     3661 gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct
     3721 gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct
     3781 gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa
     3841 ccggctgtcc gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc
     3901 catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc
     3961 ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct
     4021 gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact
     4081 ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt
     4141 ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg
     4201 ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca
     4261 gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc
     4321 cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg
     4381 cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa
     4441 ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac
     4501 cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga
     4561 gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca
     4621 agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct
     4681 gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta
     4741 cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa
     4801 gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag
     4861 aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa
     4921 aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa
     4981 gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa
     5041 atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga
     5101 taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga
     5161 gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct
     5221 gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca
     5281 cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat
     5341 cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag
     5401 catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acaaaaggcc
     5461 ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag gaattcggca gtggagaggg
     5521 cagaggaagt ctgctaacat gcggtgacgt cgaggagaat cctggcccag tgagcaaggg
     5581 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     5641 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     5701 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     5761 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     5821 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     5881 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     5941 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     6001 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     6061 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     6121 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     6181 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     6241 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aaggaattct aactagagct
     6301 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     6361 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     6421 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     6481 agcaaggggg aggattggga agagaatagc aggcatgctg gggagcggcc gcaggaaccc
     6541 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     6601 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     6661 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     6721 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     6781 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     6841 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     6901 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     6961 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     7021 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     7081 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     7141 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     7201 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     7261 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     7321 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     7381 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     7441 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     7501 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     7561 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     7621 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     7681 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     7741 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     7801 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     7861 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     7921 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     7981 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     8041 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     8101 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     8161 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     8221 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     8281 ataaatctgg agccggtgag cgtggaagcc gcggtatcat tgcagcactg gggccagatg
     8341 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     8401 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     8461 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     8521 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     8581 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     8641 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     8701 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     8761 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     8821 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     8881 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     8941 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     9001 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     9061 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     9121 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     9181 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     9241 tggccttttg ctggcctttt gctcacatgt
//
